Temeyer KB

General

Full name : Temeyer Kevin B

First name : Kevin B

Mail : Agricultural Research Service\; U.S. Department of Agriculture\; Knipling- Bushland U.S. Livestock Insects Research Laboratory\; 2700 Fredericksburg Road\; Kerrville\; 78028

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Country : USA

Email : kevin.temeyer@ars.usda.gov

Phone : +18307920330

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References (26)

Title : Identification, Baculoviral Expression, and Biochemical Characterization of a Novel Cholinesterase of Amblyomma americanum (Acari: Ixodidae) - Temeyer_2023_Int.J.Mol.Sci_24_7681
Author(s) : Temeyer KB , Schlechte KG , Gross AD , Lohmeyer KH
Ref : Int J Mol Sci , 24 :7681 , 2023
Abstract : A cDNA encoding a novel cholinesterase (ChE, EC 3.1.1.8) from the larvae of Amblyomma americanum (Linnaeus) was identified, sequenced, and expressed in Sf21 insect cell culture using the baculoviral expression vector pBlueBac4.5/V5-His. The open reading frame (1746 nucleotides) of the cDNA encoded 581 amino acids beginning with the initiation codon. Identical cDNA sequences were amplified from the total RNA of adult tick synganglion and salivary gland, strongly suggesting expression in both tick synganglion and saliva. The recombinant enzyme (rAaChE1) was highly sensitive to eserine and BW284c51, relatively insensitive to tetraisopropyl pyrophosphoramide (iso-OMPA) and ethopropazine, and hydrolyzed butyrylthiocholine (BuTCh) 5.7 times as fast as acetylthiocholine (ATCh) at 120 microM, with calculated KM values for acetylthiocholine (ATCh) and butyrylthiocholine of 6.39 microM and 14.18 microM, respectively. The recombinant enzyme was highly sensitive to inhibition by malaoxon, paraoxon, and coroxon in either substrate. Western blots using polyclonal rabbit antibody produced by immunization with a peptide specific for rAaChE1 exhibited reactivity in salivary and synganglial extract blots, indicating the presence of AaChE1 antigenic protein. Total cholinesterase activities of synganglial or salivary gland extracts from adult ticks exhibited biochemical properties very different from the expressed rAaACh1 enzyme, evidencing the substantial presence of additional cholinesterase activities in tick synganglion and saliva. The biological function of AaChE1 remains to be elucidated, but its presence in tick saliva is suggestive of functions in hydrolysis of cholinergic substrates present in the large blood mean and potential involvement in the modulation of host immune responses to tick feeding and introduced pathogens.
ESTHER : Temeyer_2023_Int.J.Mol.Sci_24_7681
PubMedSearch : Temeyer_2023_Int.J.Mol.Sci_24_7681
PubMedID: 37175388
Gene_locus related to this paper: ambam-AaChE1

Title : Effects of essential oils on native and recombinant acetylcholinesterases of Rhipicephalus microplus - Santos_2021_Rev.Bras.Parasitol.Vet_30_e002221
Author(s) : Santos E , Bezerra W , Temeyer KB , Leon AAP , Costa-Junior LM , Soares A
Ref : Rev Bras Parasitol Vet , 30 :e002221 , 2021
Abstract : This study reports the action of essential oils (EO) from five plants on the activity of native and recombinant acetylcholinesterases (AChE) from Rhipicephalus microplus. Enzyme activity of native susceptible AChE extract (S.AChE), native resistant AChE extract (R.AChE), and recombinant enzyme (rBmAChE1) was determined. An acetylcholinesterase inhibition test was used to verify the effect of the EO on enzyme activity. EO from Eucalyptus globulus, Citrus aurantifolia, Citrus aurantium var.dulcis inhibited the activity of S.AChE and R.AChE. Oils from the two Citrus species inhibited S.AChE and R.AChE in a similar way while showing greater inhibition on R.AChE. The oil from E. globulus inhibited native AChE, but no difference was observed between the S.AChE and R.AChE; however, 71% inhibition for the rBmAChE1 was recorded. Mentha piperita oil also inhibited S.AChE and R.AChE, but there was significant inhibition at the highest concentration tested. Cymbopogon winterianus oil did not inhibit AChE. Further studies are warranted with the oils from the two Citrus species that inhibited R.AChE because of the problem with R. microplus resistant to organophosphates, which target AChE. C. winterianus oil can be used against R. microplus populations that are resistant to organophosphates because its acaricidal properties act by mechanism(s) other than AChE inhibition.
ESTHER : Santos_2021_Rev.Bras.Parasitol.Vet_30_e002221
PubMedSearch : Santos_2021_Rev.Bras.Parasitol.Vet_30_e002221
PubMedID: 34076049

Title : Association of Salivary Cholinesterase With Arthropod Vectors of Disease - Temeyer_2020_J.Med.Entomol__
Author(s) : Temeyer KB , Schlechte KG , Olafson PU , Drolet BS , Tidwell JP , Osbrink WLA , Showler AT , Gross AD , Perez de Leon AA
Ref : Journal of Medical Entomology , : , 2020
Abstract : Acetylcholinesterase (AChE) was previously reported to be present in saliva of the southern cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini), with proposed potential functions to 1) reduce acetylcholine toxicity during rapid engorgement, 2) modulate host immune responses, and 3) to influence pathogen transmission and establishment in the host. Potential modulation of host immune responses might include participation in salivary-assisted transmission and establishment of pathogens in the host as has been reported for a number of arthropod vector-borne diseases. If the hypothesis that tick salivary AChE may alter host immune responses is correct, we reasoned that similar cholinesterase activities might be present in saliva of additional arthropod vectors. Here, we report the presence of AChE-like activity in the saliva of southern cattle ticks, Rhipicephalus (Boophilus) microplus; the lone star tick, Amblyomma americanum (Linnaeus); Asian tiger mosquitoes, Aedes albopictus (Skuse); sand flies, Phlebotomus papatasi (Scopoli); and biting midges, Culicoides sonorensis Wirth and Jones. Salivary AChE-like activity was not detected for horn flies Haematobia irritans (L.), stable flies Stomoxys calcitrans (L.), and house flies Musca domestica L. Salivary cholinesterase (ChE) activities of arthropod vectors of disease-causing agents exhibited various Michaelis-Menten KM values that were each lower than the KM value of bovine serum AChE. A lower KM value is indicative of higher affinity for substrate and is consistent with a hypothesized role in localized depletion of host tissue acetylcholine potentially modulating host immune responses at the arthropod bite site that may favor ectoparasite blood-feeding and alter host defensive responses against pathogen transmission and establishment.
ESTHER : Temeyer_2020_J.Med.Entomol__
PubMedSearch : Temeyer_2020_J.Med.Entomol__
PubMedID: 32459332

Title : Terpenes on Rhipicephalus (Boophilus) microplus: Acaricidal activity and acetylcholinesterase inhibition - Cardoso_2020_Vet.Parasitol_280_109090
Author(s) : Cardoso ADS , Santos EGG , Lima ADS , Temeyer KB , Perez de Leon AA , Costa LMJ , Soares A
Ref : Vet Parasitol , 280 :109090 , 2020
Abstract : The Rhipicephalus (Boophilus) microplus tick is the main ectoparasite of cattle in tropical and subtropical regions worldwide. Resistance to chemical acaricides has become widespread affirming the need for new drugs to tick control. Terpenes have become a promising alternative for cattle tick control, however the mechanism of action of these compounds is still controversial. Inhibition of acetylcholinesterase (AChE) is a well established mechanism of action of organophosphate and carbamate acaricides, but the possible action of terpenes on tick AChEs has seldom been studied in resistant and sensitive strains of R. (B.) microplus. The aim of the present study was to evaluate terpene inhibition of AChE from resistant and sensitive strains of R. (B.) microplus in correlation with their acaricidal activity. Among the terpenes used in the present study, p-cymene, thymol, carvacrol, and citral displayed acaricidal activity with LC50 of 1.75, 1.54, 1.41, and 0.38mg.mL(-1) for the susceptible strain, and LC50 of 1.40, 1.81, 1.10, and 1.13mg.mL(-1) for the resistant strain. Thymol and carvacrol inhibited the AChE of the susceptible strain larvae with IC50 of 0.93 and 0.04mg.mL(-1), respectively. The IC50 exhibited by eucalyptol, carvacrol and thymol for AChE of the resistant strain larvae were 0.36, 0.28, and 0.13mg.mL(-1), respectively. This was the first study to investigate the action of terpenes on AChE from susceptible and resistant R. (B.) microplus. As not all terpenes with acaridical activity showed AChE inhibition, the participation of AChE in the acaricidal activity of terpenes needs further investigation.
ESTHER : Cardoso_2020_Vet.Parasitol_280_109090
PubMedSearch : Cardoso_2020_Vet.Parasitol_280_109090
PubMedID: 32208306

Title : Baculoviral Expression of Presumptive OP-Resistance Mutations in BmAChE1 of Rhipicephalus (Boophilus) microplus (Ixodida: Ixodidae) and Biochemical Resistance to OP Inhibition - Temeyer_2019_J.Med.Entomol_56_1318
Author(s) : Temeyer KB , Schlechte KG , McDonough WP
Ref : Journal of Medical Entomology , 56 :1318 , 2019
Abstract : The southern cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini), transmits bovine babesiosis and anaplasmosis, and is endemic to Mexico, Latin and South America. Rhipicephalus (B.) microplus infestations within the United States are a continuing threat to U.S. cattle producers. An importation barrier between Texas and Mexico keeps the ticks from re-entering the United States. All cattle imported into the United States are dipped in an organophosphate (OP) acaricide and hand inspected for presence of ticks. Tick resistance has developed to most available acaricides, including coumaphos, the OP used in the cattle dip vats. OP-resistance can result from one or more mutations in the gene encoding the enzyme, acetylcholinesterase (AChE), resulting in production of an altered AChE resistant to OP inhibition. Previous research reported a large number of BmAChE1 mutations associated with OP resistance. We report baculovirus expression of recombinant tick BmAChE1 (rBmAChE) enzymes containing a single resistance-associated mutation, to assess their contribution to OP inhibition resistance. Surprisingly, of the naturally occurring BmAChE1 resistance-associated mutations, only D188G resulted in markedly reduced sensitivity to OP-inhibition suggesting that OP-insensitivity in BmAChE1 may result from the D188G mutation, or may possibly result from multiple mutations, each contributing a small decrease in OP sensitivity. Furthermore, an OP-insensitivity mutation (G119S) found in mosquitoes was expressed in rBmAChE1, resulting in 500-2000-fold decreased sensitivity to OP inhibition. Recombinant BmAChE1 with the G119S mutation demonstrated the lack of any structural prohibition to broad and high-level OP-insensitivity, suggesting potential increases in tick OP-resistance that would threaten the U.S. importation barrier to ticks.
ESTHER : Temeyer_2019_J.Med.Entomol_56_1318
PubMedSearch : Temeyer_2019_J.Med.Entomol_56_1318
PubMedID: 31102447
Gene_locus related to this paper: boomi-ACHE1

Title : Molecular biology of tick acetylcholinesterases - Temeyer_2018_Front.Biosci.(Landmark.Ed)_23_1320
Author(s) : Temeyer KB
Ref : Front Biosci (Landmark Ed) , 23 :1320 , 2018
Abstract : Ticks vector many pathogens with major health and economic impacts and have developed resistance to most acaricides used for tick control. Organophosphate (OP) acaricides target acetylcholinesterase (AChE) critical to tick central nervous system function. Mutations producing tick AChEs resistant to OPs were characterized; but tick OP-resistance is not fully elucidated, due to remarkable complexity of tick cholinergic systems. Three paralogous tick AChEs exhibiting differences in primary structure and biochemical kinetics are encoded by amplified genes with developmentally regulated expression. Gene silencing data suggest tick AChEs are functional complements in vivo, and transcriptomic and genomic data suggest existence of additional tick AChEs. Cholinergic systems are crucial in neural transmission and are also regulators of vertebrate immune function. Ticks exhibit prolonged intimate host contact, suggesting adaptive functions for tick cholinergic system complexity. AChE was recently reported in tick saliva and a role in manipulation of host immune responses was hypothesized. Physiological roles and genetic control of multiple tick AChEs requires further elucidation and may provide unique opportunities to understand and manipulate cholinergic involvement in biological systems.
ESTHER : Temeyer_2018_Front.Biosci.(Landmark.Ed)_23_1320
PubMedSearch : Temeyer_2018_Front.Biosci.(Landmark.Ed)_23_1320
PubMedID: 28930602
Gene_locus related to this paper: boomi-ACHE1 , boomi-ACHE2 , boomi-ACHE3

Title : Tick Salivary Cholinesterase: A Probable Immunomodulator of Host-parasite Interactions - Temeyer_2016_J.Med.Entomol_53_500
Author(s) : Temeyer KB , Tuckow AP
Ref : Journal of Medical Entomology , 53 :500 , 2016
Abstract : The southern cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini), is the most economically important cattle ectoparasite in the world. Rhipicephalus microplus and Rhipicephalus annulatus (Say) continue to threaten U.S. cattle producers despite eradication and an importation barrier based on inspection, dipping of imported cattle in organophosphate (OP) acaricide, and quarantine of infested premises. OP acaricides inhibit acetylcholinesterase (AChE), essential to tick central nervous system function. Unlike vertebrates, ticks possess at least three genes encoding AChEs, differing in amino acid sequence and biochemical properties. Genomic analyses of R. microplus and the related tick, Ixodes scapularis, suggest that ticks contain many genes encoding different AChEs. This work is the first report of a salivary cholinesterase (ChE) activity in R. microplus, and discusses complexity of the cholinergic system in ticks and significance of tick salivary ChE at the tick-host interface. It further provides three hypotheses that the salivary ChE plausibly functions 1) to reduce presence of potentially toxic acetylcholine present in the large bloodmeal imbibed during rapid engorgement, 2) to modulate the immune response (innate and/or acquired) of the host to tick antigens, and 3) to influence transmission and establishment of pathogens within the host animal. Ticks are vectors for a greater number and variety of pathogens than any other parasite, and are second only to mosquitoes (owing to malaria) as vectors of serious human disease. Saliva-assisted transmission (SAT) of pathogens is well-known; however, the salivary components participating in the SAT process remain to be elucidated.
ESTHER : Temeyer_2016_J.Med.Entomol_53_500
PubMedSearch : Temeyer_2016_J.Med.Entomol_53_500
PubMedID: 26794231

Title : Acetylcholinesterase of the sand fly, Phlebotomus papatasi (Scopoli): construction, expression and biochemical properties of the G119S orthologous mutant - Temeyer_2014_Parasit.Vectors_7_577
Author(s) : Temeyer KB , Tong F , Totrov MM , Tuckow AP , Chen QH , Carlier PR , Perez de Leon AA , Bloomquist JR
Ref : Parasit Vectors , 7 :577 , 2014
Abstract : Background Phlebotomus papatasi vectors zoonotic cutaneous leishmaniasis. Previous expression of recombinant P. papatasi acetylcholinesterase (PpAChE1) revealed 85% amino acid sequence identity to mosquito AChE and identified synthetic carbamates that effectively inhibited PpAChE1 with improved specificity for arthropod AChEs compared to mammalian AChEs. We hypothesized that the G119S mutation causing high level resistance to organophosphate insecticides in mosquitoes may occur in PpAChE1 and may reduce sensitivity to inhibition. We report construction, expression, and biochemical properties of rPpAChE1 containing the G119S orthologous mutation.MethodsTargeted mutagenesis introduced the G119S orthologous substitution in PpAChE1 cDNA. Recombinant PpAChE1 enzymes containing or lacking the G119S mutation were expressed in the baculoviral system. Biochemical assays were conducted to determine altered catalytic properties and inhibitor sensitivity resulting from the G119S substitution. A molecular homology model was constructed to examine the modeled structural interference with docking of inhibitors of different classes. Genetic tests were conducted to determine if the G119S orthologous codon existed in polymorphic form in a laboratory colony of P. papatasi.ResultsRecombinant PpAChE1 containing the G119 substitution exhibited altered biochemical properties, and reduced inhibition by compounds that bind to the acylation site on the enzyme (with the exception of eserine). Less resistance was directed against bivalent or peripheral site inhibitors, in good agreement with modeled inhibitor docking. Eserine appeared to be a special case capable of inhibition in the absence of covalent binding at the acylation site. Genetic tests did not detect the G119S mutation in a laboratory colony of P. papatasi but did reveal that the G119S codon existed in polymorphic form (GGA + GGC).ConclusionsThe finding of G119S codon polymorphism in a laboratory colony of P. papatasi suggests that a single nucleotide transversion (GGC inverted question mark AGC) may readily occur, causing rapid development of resistance to organophosphate and phenyl-substituted carbamate insecticides under strong selection. Careful management of pesticide use in IPM programs is important to prevent or mitigate development and fixation of the G119S mutation in susceptible pest populations. Availability of recombinant AChEs enables identification of novel inhibitory ligands with improved efficacy and specificity for AChEs of arthropod pests.
ESTHER : Temeyer_2014_Parasit.Vectors_7_577
PubMedSearch : Temeyer_2014_Parasit.Vectors_7_577
PubMedID: 25491113
Gene_locus related to this paper: phlpp-m1f887

Title : Inhibitor profile of (n)-tacrines -methylcarbamates on acetylcholinesterase from Rhipicephalus (Boophilus) microplus and Phlebotomus papatasi - Swale_2013_Pestic.Biochem.Physiol_106_
Author(s) : Swale DR , Tong F , Temeyer KB , Li A , Lam PC , Totrov MM , Carlier PR , Perez de Leon AA , Bloomquist JR
Ref : Pesticide Biochemistry and Physiology , 106 : , 2013
Abstract : The cattle tick, Rhipicephalus (Boophilus) microplus (Bm), and the sand fly, Phlebotomus papatasi (Pp), are disease vectors to cattle and humans, respectively. The purpose of this study was to characterize the inhibitor profile of acetylcholinesterases from Bm (BmAChE1) and Pp (PpAChE) compared to human and bovine AChE, in order to identify divergent pharmacology that might lead to selective inhibitors. Results indicate that BmAChE has low sensitivity (IC50 = 200 muM) toward tacrine, a monovalent catalytic site inhibitor with sub micromolar blocking potency in all previous species tested. Similarly, a series of bis(n)-tacrine dimer series, bivalent inhibitors and peripheral site AChE inhibitors possess poor potency toward BmAChE. Molecular homology models suggest the rBmAChE enzyme possesses a W384F orthologous substitution near the catalytic site, where the larger tryptophan side chain obstructs the access of larger ligands to the active site, but functional analysis of this mutation suggests it only partially explains the low sensitivity to tacrine. In addition, BmAChE1 and PpAChE have low nanomolar sensitivity to some experimental carbamate anticholinesterases originally designed for control of the malaria mosquito, Anopheles gambiae. One experimental compound, 2-((2-ethylbutyl)thio)phenyl methylcarbamate, possesses >300-fold selectivity for BmAChE1 and PpAChE over human AChE, and a mouse oral LD50 of >1500 mg/kg, thus providing an excellent new lead for vector control.
ESTHER : Swale_2013_Pestic.Biochem.Physiol_106_
PubMedSearch : Swale_2013_Pestic.Biochem.Physiol_106_
PubMedID: 24187393
Gene_locus related to this paper: boomi-ACHE1 , boomi-ACHE2 , boomi-ACHE3

Title : Acetylcholinesterase of Rhipicephalus (Boophilus) microplus and Phlebotomus papatasi: Gene identification, expression, and biochemical properties of recombinant proteins - Temeyer_2013_Pestic.Biochem.Physiol_106_118
Author(s) : Temeyer KB , Olafson PU , Brake DK , Tuckow AP , Li AY , Perez de Leon AA
Ref : Pesticide Biochemistry and Physiology , 106 :118 , 2013
Abstract : The southern cattle tick Rhipicephalus (Boophilus) microplus (Bm) is a vector of bovine babesiosis and anaplasmosis. Tick resistance to organophosphate (OP) acaricide involves acetylcholinesterase (AChE) insensitivity to OP and metabolic detoxification. Sequencing and in vitro expression of Bm genes encoding AChE allowed biochemical characterization of three BmAChEs expressed in tick synganglion. rBmAChE1, rBmAChE2 and rBmAChE3 exhibited substrate preference for acetylthiocholine, high substrate inhibition and sensitivity to AChE-specific inhibitors. OP-insensitivity mutations were demonstrated in rBmAChE1 and rBmAChE3. Gene silencing suggested functional complementation of BmAChEs in vivo. BmAChE genes were amplified in copy number and multiple transcript polymorphisms were expressed in individual tick synganglia for each of the BmAChEs, suggesting allelic diversity within individuals. Studies also identified a gene encoding AChE of the sand fly, Phlebotomus papatasi, a vector of leishmaniasis in humans and animals. Expression of recombinant P. papatasi AChE (rPpAChE) enabled biochemical characterization and identification of effective inhibitors that selectively target rPpAChE.
ESTHER : Temeyer_2013_Pestic.Biochem.Physiol_106_118
PubMedSearch : Temeyer_2013_Pestic.Biochem.Physiol_106_118
PubMedID:
Gene_locus related to this paper: boomi-ACHE1 , boomi-ACHE2 , boomi-ACHE3

Title : Acetylcholinesterase of the sand fly, Phlebotomus papatasi (Scopoli): cDNA sequence, baculovirus expression, and biochemical properties - Temeyer_2013_Parasit.Vectors_6_31
Author(s) : Temeyer KB , Brake DK , Tuckow AP , Li AY , Perez de Leon AA
Ref : Parasit Vectors , 6 :31 , 2013
Abstract : ABSTRACT: BACKGROUND: Millions of people and domestic animals around the world are affected by leishmaniasis, a disease caused by various species of flagellated protozoans in the genus Leishmania that are transmitted by several sand fly species. Insecticides are widely used for sand fly population control to try to reduce or interrupt Leishmania transmission. Zoonotic cutaneous leishmaniasis caused by L. major is vectored mainly by Phlebotomus papatasi (Scopoli) in Asia and Africa. Organophosphates comprise a class of insecticides used for sand fly control, which act through the inhibition of acetylcholinesterase (AChE) in the central nervous system. Point mutations producing an altered, insensitive AChE are a major mechanism of organophosphate resistance in insects and preliminary evidence for organophosphate-insensitive AChE has been reported in sand flies. This report describes the identification of complementary DNA for an AChE in P. papatasi and the biochemical characterization of recombinant P. papatasi AChE.
METHODS: A P. papatasi Israeli strain laboratory colony was utilized to prepare total RNA utilized as template for RT-PCR amplification and sequencing of cDNA encoding acetylcholinesterase 1 using gene specific primers and 3'-5'RACE. The cDNA was cloned into pBlueBac4.5/V5-His TOPO, and expressed by baculovirus in Sf21 insect cells in serum-free medium. Recombinant P. papatasi acetylcholinesterase was biochemically characterized using a modified Ellman's assay in microplates.
RESULTS: A 2309 nucleotide sequence of PpAChE1 cDNA [GenBank: JQ922267] of P. papatasi from a laboratory colony susceptible to insecticides is reported with 73-83% nucleotide identity to acetylcholinesterase mRNA sequences of Culex tritaeniorhynchus and Lutzomyia longipalpis, respectively. The P. papatasi cDNA ORF encoded a 710-amino acid protein [GenBank: AFP20868] exhibiting 85% amino acid identity with acetylcholinesterases of Cx. pipiens, Aedes aegypti, and 92% amino acid identity for L. longipalpis.Recombinant P. papatasi AChE1 was expressed in the baculovirus system and characterized as an insect acetylcholinesterase with substrate preference for acetylthiocholine and inhibition at high substrate concentration. Enzyme activity was strongly inhibited by eserine, BW284c51, malaoxon, and paraoxon, and was insensitive to the butyrylcholinesterase inhibitors ethopropazine and iso-OMPA.
CONCLUSIONS: Results presented here enable the screening and identification of PpAChE mutations resulting in the genotype for insensitive PpAChE. Use of the recombinant P. papatasi AChE1 will facilitate rapid in vitro screening to identify novel PpAChE inhibitors, and comparative studies on biochemical kinetics of inhibition.
ESTHER : Temeyer_2013_Parasit.Vectors_6_31
PubMedSearch : Temeyer_2013_Parasit.Vectors_6_31
PubMedID: 23379291
Gene_locus related to this paper: phlpp-m1f887

Title : Discovery of microRNAs of the stable fly (Diptera: Muscidae) by High-throughput sequencing - Tuckow_2013_J.Med.Entomol_50_925
Author(s) : Tuckow AP , Temeyer KB , Olafson PU , Perez de Leon AA
Ref : Journal of Medical Entomology , 50 :925 , 2013
Abstract : The stable fly, Stomoxys calcitrans (L.), is a serious ectoparasite affecting animal production and health of both animals and humans. Stable fly control relies largely on chemical insecticides; however, the development of insecticide resistance as well as environmental considerations requires continued discovery research to develop novel control technologies. MicroRNAs (miRNAs) are a class of short noncoding RNAs that have been shown to be important regulators of gene expression across a wide variety of organisms, and may provide an innovative approach with regard to development of safer more targeted control technologies. The current study reports discovery ad initial comparative analysis of 88 presumptive miRNA sequences from the stable fly, obtained using high-throughput sequencing of small RNAs. The majority of stable fly miRNAs were 22-23 nt in length. Many miRNAs were arthropod specific, and several mature miRNA sequences showed greater sequence identity to miRNAs from other blood-feeding dipterans such as mosquitoes rather than to Drosophilids. This initial step in characterizing the stable fly microRNAome provides a basis for further analyses of life stage-specific and tissue-specific expression to elucidate their functional roles in stable fly biology.
ESTHER : Tuckow_2013_J.Med.Entomol_50_925
PubMedSearch : Tuckow_2013_J.Med.Entomol_50_925
PubMedID: 23926794

Title : Acetylcholinesterases of blood-feeding flies and ticks - Temeyer_2013_Chem.Biol.Interact_203_319
Author(s) : Temeyer KB , Tuckow AP , Brake DK , Li AY , Perez de Leon AA
Ref : Chemico-Biological Interactions , 203 :319 , 2013
Abstract : Acetylcholinesterase (AChE) is the biochemical target of organophosphate (OP) and carbamate pesticides for invertebrates, vertebrate nerve agents, and AChE inhibitors used to reduce effects of Alzheimer's disease. Organophosphate pesticides (OPs) are widely used to control blood-feeding arthropods, including biting flies and ticks. However, resistance to OPs in pests affecting animal and human health has compromised control efficacy. OP resistance often results from mutations producing an OP-insensitive AChE. Our studies have demonstrated production of OP-insensitive AChEs in biting flies and ticks. Complementary DNA (cDNA) sequences encoding AChEs were obtained for the horn fly, stable fly, sand fly, and the southern cattle tick. The availability of cDNA sequences enables the identification of mutations, expression and characterization of recombinant proteins, gene silencing for functional studies, as well as in vitro screening of novel inhibitors. The southern cattle tick expresses at least three different genes encoding AChE in their synganglion, i.e. brain. Gene amplification for each of the three known cattle tick AChE genes and expression of multiple alleles for each gene may reduce fitness cost associated with OP-resistance. AChE hydrolyzes the neurotransmitter, acetylcholine, but may have additional roles in physiology and development. The three cattle tick AChEs possess significantly different biochemical properties, and are expressed in neural and non-neural tissues, which suggest separation of structure and function. The remarkable complexity of AChEs in ticks suggested by combining genomic data from Ixodes scapularis with our genetic and biochemical data from Rhipicephalus microplus is suggestive of previously unknown gene duplication and diversification. Comparative studies between invertebrate and vertebrate AChEs could enhance our understanding of structure-activity relationships. Research with ticks as a model system offers the opportunity to elucidate structure-activity relationships for AChE that are important for advances in targeted pest control, as well as potential applications for medicine and biosecurity.
ESTHER : Temeyer_2013_Chem.Biol.Interact_203_319
PubMedSearch : Temeyer_2013_Chem.Biol.Interact_203_319
PubMedID: 23036311

Title : Sequence polymorphism in acetylcholinesterase transcripts and genotyping survey of BmAchE1 in laboratory and Mexican strains of Rhipicephalus (Boophilus) microplus - Temeyer_2012_J.Med.Entomol_49_555
Author(s) : Temeyer KB , Olafson PU , Pruett JH
Ref : Journal of Medical Entomology , 49 :555 , 2012
Abstract : ABSTRACT Acetylcholinesterase cDNAs, BmAChE1, BmAChE2, and BmAChE3 of Rhipicephalus (Boophilus) microplus (Canestrini) were sequenced and found to exhibit significant polymorphism. A portion of the predicted amino acid substitutions in BmAChE1, BmAChE2, and BmAChE3 were found predominantly in organophosphate-resistant strains, but most did not correlate with resistant status. Multiple transcripts were observed from individual ticks, suggesting possible gene duplication or alternative splicing to produce more than two transcripts per individual. BmAChE1 transcript polymorphisms associating with organophosphate-resistant status in laboratory strains were surveyed in laboratory and Mexican strains of R. microplus by sequencing BmAChE1 genomic DNA. Quantitative real-time polymerase chain reaction was used to determine copy numbers of BmAChE1 (eight copies/haploid genome), BmAChE2 (16 copies/haploid genome), and BmAChE3 (four copies/haploid genome). Presence of at least three highly polymorphic amplified genes expressing AChE in tick synganglion suggested that ticks maintain a large and diverse assortment of AChE alleles available for rapid recombination and selection, which potentially reduces fitness costs associated with individual mutations. Elevated copy numbers for each of the BmAChEs may also explain previous failures to identify mutations resulting in insensitivity to organophosphates. It is clear that development of phenotypic resistance to organophosphates is highly complex and may be multigenic in character.
ESTHER : Temeyer_2012_J.Med.Entomol_49_555
PubMedSearch : Temeyer_2012_J.Med.Entomol_49_555
PubMedID: 22679862
Gene_locus related to this paper: boomi-ACHE1 , boomi-ACHE2 , boomi-ACHE3

Title : Acetylcholinesterase of Haematobia irritans (Diptera: Muscidae): baculovirus expression, biochemical properties, and organophosphate insensitivity of the G262A mutant - Temeyer_2012_J.Med.Entomol_49_589
Author(s) : Temeyer KB , Brake DK , Schlechte KG
Ref : Journal of Medical Entomology , 49 :589 , 2012
Abstract : This study reports the baculovirus expression and biochemical characterization of recombinant acetylcholinesterase from Haematobia irritans (L.) (rHiAChE) and the effect of the previously described G262A mutation on enzyme activity and sensitivity to selected organophosphates. The rHiAChE was confirmed to be an insect AChE2-type enzyme with substrate preference for acetylthiocholine (Km 31.3 microM) over butyrylthiocholine (Km 63.4 microM) and inhibition at high substrate concentration. Enzyme activity was strongly inhibited by eserine (2.3 x 10(-10) M), BW284c51 (3.4 x 10(-8) M), malaoxon (3.6 x 10(-9) M), and paraoxon (1.8 x 10(-7) M), and was less sensitive to the butyrylcholinesterase inhibitors ethopropazine (1.1 x 10(-6) M) and iso-OMPA (4.1 x 10(-4) M). rHiAChE containing the G262A substitution exhibited decreased substrate affinity for both acetylthiocholine (Km 40.9 microM) and butyrylthiocholine (Km 96.3 microM), and exhibited eight-fold decreased sensitivity to paraoxon, and approximately 1.5- to 3-fold decreased sensitivity to other inhibitors. The biochemical kinetics are consistent with previously reported bioassay analysis, suggesting that the G262A mutation contributes to, but is not solely responsible for observed phenotypic resistance to diazinon or other organophosphates.
ESTHER : Temeyer_2012_J.Med.Entomol_49_589
PubMedSearch : Temeyer_2012_J.Med.Entomol_49_589
PubMedID: 22679866
Gene_locus related to this paper: haeir-ACHE

Title : Acetylcholinesterase of Stomoxys calcitrans (L.) (Diptera: Muscidae): cDNA sequence, baculovirus expression, and biochemical properties - Temeyer_2012_Vet.Parasitol_184_92
Author(s) : Temeyer KB , Chen AC
Ref : Vet Parasitol , 184 :92 , 2012
Abstract : A 2193-nucleotide cDNA encoding acetylcholinesterase (AChE) of the stable fly, Stomoxys calcitrans (L.) was sequenced and expressed in the baculovirus system. The open reading frame encoded a 91 amino acid secretion signal peptide and a 613 amino acid mature protein with 96% and 94% identity to the AChEs of Haematobia irritans (L.) and Musca domestica (L.), respectively. Structural characteristics of M. domestica, H. irritans, and Drosophila melanogaster AChEs were conserved in the S. calcitrans AChE. The recombinant enzyme was inhibited by eserine, coroxon, and paraoxon and exhibited K(m) values of 63.9muM for acetylthiocholine and 96.7muM for butyrylthiocholine, confirming its biochemical identity as an acetylcholinesterase (EC 3.1.1.7). These data will enable rapid identification and assay for mutations that reduce AChE sensitivity to organophosphate (OP) pesticides, potentially aiding resistance management efforts to prevent fixation of the mutations in pest populations.
ESTHER : Temeyer_2012_Vet.Parasitol_184_92
PubMedSearch : Temeyer_2012_Vet.Parasitol_184_92
PubMedID: 21872994
Gene_locus related to this paper: stoca-ACHE

Title : Genetic factors potentially reducing fitness cost of organophosphate insensitive acetylcholinesterase(s) in Rhipicephalus (Boophilus) microplus - Temeyer_2011_Zoosymposia_6_260
Author(s) : Temeyer KB , Tijerina MA , Davey RB , Olafson PU
Ref : Zoosymposia , 6 :260 , 2011
Abstract : Acaricidal activity of organophosphate (OP) and carbamate acaricides is believed to result from inhibition of acetylcholinesterase (AChE). Previous studies in Rhipicephalus (Boophilus) microplus demonstrated the presence of three presumptive AChE genes (BmAChEs). Biochemical characterization of recombinant BmAChE proteins expressed in the baculovirus system demonstrated that each of the three R. (B.) microplus rBmAChEs have enzymatic properties consistent with designation as functional acetylcholinesterases. Complementary DNAs (cDNAs) for each of the three BmAChEs were cloned and sequenced from individual adult tick synganglia excised from OP-susceptible and OPresistant strains. The data revealed the presence of multiple transcript sequences within individual ticks for each of the BmAChEs, suggesting alternative mRNA splicing or expression of multiple alleles for each of the BmAChE genes. Quantitative real-time PCR provided evidence of possible gene duplication or amplification for each of the BmAChE genes, and direct sequencing of genomic DNA provided evidence of structural BmAChE gene diversity with respect to presence or absence of introns, as well as the presence or absence of sequence polymorphisms. Baculovirus expression of rBmAChE1 and rBmAChE3 proteins containing some of the predicted amino acid sequence polymorphisms resulted in production of OP-insensitive AChE, demonstrating that at least some OP-resistant individuals contain mutations that reduce OP-inhibition for at least two of the three known BmAChEs. RNA interference was utilized to silence in vivo expression of the BmAChE genes in adult ticks, resulting in tick mortality if all three BmAChEs were silenced simultaneously, strongly suggesting that the BmAChE proteins functionally complement one another in vivo. Together, the results presented provide strong evidence that OP-resistance in R. microplus is at least partially mediated by a combination of the expression of multiple genes encoding acetylcholinesterase, mutations in BmAChEs resulting in OP-insensitivity, gene duplication, and maintenance of allelic diversity, including both OP-sensitive and OPinsensitive alleles within individual ticks. The authors propose that these factors may mitigate fitness costs that might otherwise result from BmAChE mutations, and demonstrate the extreme complexity of OP-resistance in R. (B.) microplus. It is hoped that elucidation of the complex interactions among the multiple BmAChEs and their physiological roles may enable development of new opportunities for tick control.
ESTHER : Temeyer_2011_Zoosymposia_6_260
PubMedSearch : Temeyer_2011_Zoosymposia_6_260
PubMedID:
Gene_locus related to this paper: boomi-ACHE1 , boomi-ACHE2 , boomi-ACHE3

Title : Baculovirus expression, biochemical characterization and organophosphate sensitivity of rBmAChE1, rBmAChE2, and rBmAChE3 of Rhipicephalus (Boophilus) microplus - Temeyer_2010_Vet.Parasitol_172_114
Author(s) : Temeyer KB , Pruett JH , Olafson PU
Ref : Vet Parasitol , 172 :114 , 2010
Abstract : Rhipicephalus (Boophilus) microplus cDNAs, BmAChE1, BmAChE2, and BmAChE3, were previously identified as presumptively encoding acetylcholinesterases (AChEs), but biochemical identity was confirmed only for recombinant BmAChE3. In the present study, four recombinant BmAChE1 constructs and single recombinant constructs of BmAChE2 and BmAChE3 were expressed in baculovirus. Biochemical characterization of the recombinant proteins supports classification of rBmAChE1, rBmAChE2, and rBmAChE3 as AChEs (E.C.3.1.1.7), as evidenced by (i) substrate preference for acetylthiocholine, (ii) inhibition by eserine, BW284c51, and the organophosphates (OPs) malaoxon and paraoxon, (iii) insensitivity to iso-OMPA, and (iv) rapid hydrolysis of acetyl-beta-methyl-thiocholine. Unlike reports for insect AChEs, we did not observe substrate inhibition of activity at acetylthiocholine concentrations as high as 40 mM, however, product inhibition was apparent at 10-100 microM choline in agreement with properties reported for the catalytic domain of Anopheles gambiae acetylcholinesterase-1. Substrate affinity and V(max) values were highest for rBmAChE1 proteins, and one rBmAChE1 enzyme (Tx11, derived from the OP-resistant strain Tuxpan), was insensitive to paraoxon and exhibited a greatly reduced V(max) near that of rBmAChE2. To date, recombinant BmAChE1 and BmAChE3 enzymes with reduced sensitivity to OP-inhibition have been cloned and expressed from OP-resistant strains. The presence of at least three genes expressing AChEs in R. (B.) microplus, at least two of which contain mutations expressed as OP-insensitive enzymes, strongly suggests that phenotypic resistance to OPs may be complex and multigenic in character.
ESTHER : Temeyer_2010_Vet.Parasitol_172_114
PubMedSearch : Temeyer_2010_Vet.Parasitol_172_114
PubMedID: 20451328
Gene_locus related to this paper: boomi-ACHE1 , boomi-ACHE2 , boomi-ACHE3

Title : A survey of Rhipicephalus microplus populations for mutations associated with pyrethroid resistance - Chen_2009_J.Econ.Entomol_102_373
Author(s) : Chen AC , He H , Temeyer KB , Jones S , Green P , Barker SC
Ref : J Econ Entomol , 102 :373 , 2009
Abstract : Mutations associated with pyrethroid resistance were found in Mexican strains of Rhipicephalus microplus (Canestrini). A mutation in the sodium channel gene was reported in strains highly resistant to permethrin and another mutation in an esterase gene in a strain that shows moderate resistance to the same pesticide. Methods based on the melting temperature difference of amplified allele-specific DNA fragments were developed that can detect these mutations rapidly in individual larvae. When these methods were applied to ticks from various strains of R. microplus from Australia, neither of these mutations could be demonstrated. Different resistance mechanisms have apparently developed independently between Australian and Mexican strains of R. microplus.
ESTHER : Chen_2009_J.Econ.Entomol_102_373
PubMedSearch : Chen_2009_J.Econ.Entomol_102_373
PubMedID: 19253657

Title : Genotyping mutations in BmAChE3: A survey of organophosphate-resistant and -susceptible strains of Rhipicephalus (Boophilus) microplus - Temeyer_2009_J.Med.Entomol_46_1355
Author(s) : Temeyer KB , Olafson PU , Miller RJ
Ref : Journal of Medical Entomology , 46 :1355 , 2009
Abstract : Mutations I48L, I54V, R86Q, V137I, I492M, and T548A were identified previously in BmAChE3, a gene encoding acetylcholinesterase, from the organophosphate (OP) acaricide-resistant San Rommn strain of Rhipicephalus (Boophilus) microplus. Recombinant BmAChE3 acetylcholinesterase containing the R86Q mutation was shown to exhibit nearly 20-fold reduction in the rate of phosphorylation by paraoxon relative to the wild-type sequence. In addition, the R86Q mutation was present in resistant laboratory strains at elevated frequency compared with OP-susceptible strains but was insufficient to alone generate the OP-resistant phenotype (J. Med. Entomol. 44: 1013-1018). Here, we developed assays to genotype the remaining five mutations and evaluated frequency of all six BmAChE3 mutations in individual R. microplus ticks from laboratory and Mexican field-collected strains. We found a substantial number of individuals in known OP-susceptible strains that seemed to be homozygous for each of the mutations surveyed, the exception being I48L, which was infrequent in all strains, leading us to conclude that none of the mutations alone were responsible for generation of phenotypic resistance to OP acaricide.
ESTHER : Temeyer_2009_J.Med.Entomol_46_1355
PubMedSearch : Temeyer_2009_J.Med.Entomol_46_1355
PubMedID: 19960680
Gene_locus related to this paper: boomi-ACHE3

Title : Acetylcholinesterase mutation in diazinon-resistant Haematobia irritans (L.) (Diptera: Muscidae) - Temeyer_2008_Vet.Parasitol_154_300
Author(s) : Temeyer KB , Li AY , Lohmeyer KH , Chen AC , Olafson PU , Sanson DW , Foil LD
Ref : Vet Parasitol , 154 :300 , 2008
Abstract : Acetylcholinesterase (AChE) cDNA from individual field-collected diazinon-resistant horn flies was amplified by RT-PCR. Sequencing of the amplification products revealed that 8/12 of the diazinon-resistant horn flies contained a point mutation previously associated with resistance to organophosphates in house flies and Drosophila, strongly suggesting that this cDNA encodes the AChE that is the target site for organophosphate (OP) pesticide. The point mutation (G262A) resulted in a shift from glycine to alanine in the mature HiAChE amino acid sequence at position 262. Allele-specific PCR and RLFP assays were developed to diagnose the presence or absence of the G262A mutation in individual flies. Use of the allele-specific assays each demonstrated the presence of the G262A mutation in 10 of 12 individual field-collected flies, demonstrating higher sensitivity than direct sequencing of RT-PCR amplification products. The G262A mutation was found in additional fly populations previously characterized as OP-resistant, further supporting that this AChE is the target site for OP pesticide. The allele-specific assay is a useful tool for quantitative assay of the resistance allele in horn fly populations.
ESTHER : Temeyer_2008_Vet.Parasitol_154_300
PubMedSearch : Temeyer_2008_Vet.Parasitol_154_300
PubMedID: 18472339
Gene_locus related to this paper: haeir-ACHE

Title : R86Q, a mutation in BmAChE3 yielding a Rhipicephalus microplus organophosphate-insensitive acetylcholinesterase - Temeyer_2007_J.Med.Entomol_44_1013
Author(s) : Temeyer KB , Pruett JH , Olafson PU , Chen AC
Ref : Journal of Medical Entomology , 44 :1013 , 2007
Abstract : Mutations were identified in the cDNA sequence encoding the acetylcholinesterase BmAChE3 in strains of Rhipicephalus (Boophilus) microplus (Canestrini) resistant or susceptible to organophosphate (OP) acaricide. The mutation that occurred most frequently in the OP-resistant San Roman strain resulted in a substitution of glutamine (Q) for arginine (R) at position 86 in BmAChE3 (position 66 in mature BmAChE). Clones containing the mutant and wild-type cDNA sequences were expressed in the baculovirus system. Enzyme kinetics of recombinant BmAChE3 containing or lacking the R86Q mutation demonstrated that the R86Q mutation increased substrate affinity and conferred insensitivity to paraoxon inhibition. This is the first demonstration of a mutation in a gene encoding an ixodid acetylcholinesterase resulting in OP insensitivity. A restriction fragment length polymorphism assay was developed and used to diagnose the frequency of the R86Q mutation in BmAChE3 genomic DNA from seven laboratory-colonized strains. Use of the R86Q diagnostic assay detected an increased frequency of the R86Q mutation in OP-resistant tick strains compared with that of OP-susceptible strains; however, the R86Q mutation was also present in OP-susceptible strains at unexpectedly high frequency. Because the R86Q mutation generates an OP-resistant enzyme in vitro and it is present at an elevated frequency in laboratory strains selected for OP resistance, we conclude that the data are consistent with a potential role for BmAChE3 in development of OP resistance; however, because the R86Q mutation has a high frequency in susceptible strains, the R86Q mutation alone is insufficient to generate the OP-resistant phenotype at the organismal level. There are likely to be additional mutations in BmAChE3, mutations in additional acetylcholinesterase genes, or additional resistance mechanisms (e.g., oxidative metabolism) that contribute to expression of the OP-resistant phenotype.
ESTHER : Temeyer_2007_J.Med.Entomol_44_1013
PubMedSearch : Temeyer_2007_J.Med.Entomol_44_1013
PubMedID: 18047200
Gene_locus related to this paper: boomi-ACHE3

Title : Identification and characterization of a cDNA encoding the acetylcholinesterase of Haematobia irritans (L.) (Diptera: Muscidae) - Temeyer_2007_DNA.Seq_18_85
Author(s) : Temeyer KB , Chen AC
Ref : DNA Sequence , 18 :85 , 2007
Abstract : A 2217-nucleotide cDNA presumptively encoding acetylcholinesterase (AChE) of the horn fly, Haematobia irritans (L.) was sequenced. The open reading frame (ORF) encoded a 91 amino acid secretion signal peptide and a 613 amino acid mature protein with 95% identity and 98% similarity to the AChE of Musca domestica (L.). Structural features characteristic of the M. domestica and Drosophila melanogaster AChEs are conserved in the H. irritans AChE. The M. domestica and D. melanogaster AChEs are target sites for organophosphate inhibition as previously shown (Walsh et al. 2001. Biochem. J. 359: 175-181, Kozaki et al. 2002. Appl. Entomol. Zool. 37: 213-218), suggesting that this H. irritans AChE2 may be the target site for organophosphate.
ESTHER : Temeyer_2007_DNA.Seq_18_85
PubMedSearch : Temeyer_2007_DNA.Seq_18_85
PubMedID: 17364819
Gene_locus related to this paper: haeir-ACHE

Title : Baculovirus expression of BmAChE3, a cDNA encoding an acetylcholinesterase of Boophilus microplus (Acari: Ixodidae) - Temeyer_2006_J.Med.Entomol_43_707
Author(s) : Temeyer KB , Pruett JH , Untalan PM , Chen AC
Ref : Journal of Medical Entomology , 43 :707 , 2006
Abstract : The complete cDNA sequence encoding a Boophilus microplus (Canestrini) (Acari: Ixodidae) acetylcholinesterase (AChE3) was expressed in the baculovirus system. The recombinant AChE3 protein (rBmAChE3) was secreted as a soluble form into the cell culture medium and was identified as a functional AChE by substrate specificity and by inhibition with the AChE-specific inhibitors eserine sulfate and BW284c51. Inhibition kinetics of rBmAChE3, in the presence of the organophosphate paraoxon, revealed sensitivity comparable with that of adult, organophosphate-susceptible neural AChE. To our knowledge, this is the first report of the cloning and successful expression of a functional ixodid AChE.
ESTHER : Temeyer_2006_J.Med.Entomol_43_707
PubMedSearch : Temeyer_2006_J.Med.Entomol_43_707
PubMedID: 16892628
Gene_locus related to this paper: boomi-ACHE3

Title : Identification of a third Boophilus microplus (Acari: Ixodidae) cDNA presumptively encoding an acetylcholinesterase - Temeyer_2004_J.Med.Entomol_41_259
Author(s) : Temeyer KB , Davey RB , Chen AC
Ref : Journal of Medical Entomology , 41 :259 , 2004
Abstract : Oligodeoxynucleotide primers, based on amino acid sequences conserved in known acetylcholinesterases (AChEs), were used in reverse-transcription polymerase chain reaction (RT-PCR) with mRNA from Boophilus microplus (Canestrini) as the template. Primer walking and rapid amplification of cDNA ends (RACE) techniques were used to complete the cDNA sequence identified by RT-PCR. The complete B. microplus cDNA sequence contained an open reading frame encoding a 620 amino acid protein with a 20 amino acid signal peptide at the N-terminus targeting the protein for the secretion pathway. BLAST searches of GenBank using the presumptively encoded protein revealed highest sequence similarity to AChEs. The presumptively encoded protein was of similar size and structural properties to other identified AChEs, including the presence of the catalytic triad (Ser, Glu, His) and appropriate placement of internal cysteines to yield three internal disulfide bonds corresponding to those of known AChEs. Putative conserved domains identified the sequence as a member of the carboxylesterase family, pfam00135.8, of which AChE is a member. This cDNA therefore presumptively encodes a third transcribed AChE (AChE3) cDNA of B. microplus. Comparison of the three AChE eDNA sequences expressed in B. microplus demonstrated no discernible nucleotide sequence homology and relatively low amino acid sequence homology, strongly suggesting that they are not alleles of one another. The potential presence of multiple expressed AChEs in B. microplus suggests alternative mechanisms for development of resistance to pesticides that target AChE. The homology-based identification of a third expressed AChE in B. microplus is a surprising result and strongly implies the need for confirmation of gene identity for presumptive AChEs.
ESTHER : Temeyer_2004_J.Med.Entomol_41_259
PubMedSearch : Temeyer_2004_J.Med.Entomol_41_259
PubMedID: 15185924
Gene_locus related to this paper: boomi-ACHE3

Title : Antigenicity and immunogenicity of Hypoderma lineatum soluble proteins in the bovine host - Pruett_1988_Vet.Parasitol_29_53
Author(s) : Pruett JH , Temeyer KB , Burkett BK
Ref : Vet Parasitol , 29 :53 , 1988
Abstract : Protein species found in soluble crude extracts of Hypoderma lineatum (common cattle grub) 1st-instar larvae (HL1) were separated by non-denaturing and denaturing polyacrylamide gel electrophoresis (PAGE) and analyzed for antigenicity by Western blotting using serum from H. lineatum-infested and vaccinated cattle. All HL1 proteins resolved by non-denaturing PAGE were found to be antigenic in the infested bovine host. Treatment of the proteins with sodium dodecyl sulfate and 2-mercaptoethanol destroyed the ability of hypodermin B and the Peak 2 proteins from DEAE-ion exchange HPLC to be bound by antibody. The principal proteins, hypodermin A and hypodermin C (collagenase), appear to be the most immunogenic of the larval proteins. Although having similar amino acid composition, hypodermin A did not appear to share an antigenic epitope with the most prevalent protein, hypodermin C. These results may allow for the selection of proteins to be used in vaccine trials and studies of protective immunological mechanisms associated with acquired resistance to H. lineatum infestation in the bovine host.
ESTHER : Pruett_1988_Vet.Parasitol_29_53
PubMedSearch : Pruett_1988_Vet.Parasitol_29_53
PubMedID: 2459835