Martinez C

References (15)

Title : Biogenic metallic nanoparticles (Ag, TiO(2), Fe) as potential fungicides for agriculture: are they safe for the freshwater mussel Anodontites trapesialis? - Tesser_2022_Chemosphere_309_136664
Author(s) : Tesser ME , Guilger M , Bilesky-Jose N , Risso WE , de Lima R , Martinez C
Ref : Chemosphere , 309 :136664 , 2022
Abstract : Silver (Ag), titanium dioxide (TiO(2)), and iron (Fe) nanoparticles (NPs) synthesized using the fungus Trichoderma harzianum are effective against the agriculture pathogen Sclerotinia sclerotiorum. However, their effects should be evaluated in aquatic organisms, as agriculture practices can contaminate the aquatic environment. Thus, this work evaluated sublethal effects of acute exposure (24 h) to AgNP, TiO(2)NP and FeNP, synthesized with T. harzianum, on the Neotropical freshwater bivalve Anodontites trapesialis, considering the hypothesis that suspension-feeding bivalves are susceptible to NPs toxicity. Individuals of A. trapesialis were divided into four groups (n = 8/group): a control group, kept in water only; a group exposed to AgNP; a group exposed to TiO(2)NP; and a group exposed to FeNP. The bioaccumulation of Ag, Ti, and Fe was evaluated in the gills, hemolymph, mantle, digestive gland, and muscle (foot). Lipoperoxidation, activities of the glutathione S-transferase, catalase, and superoxide dismutase, and glycogen concentration were quantified in the gills, mantle, and digestive gland. Ions (Na(+), K(+), Cl(-), Ca(2+), and Mg(+2)) and glucose concentrations were quantified in the hemolymph. Na(+)/K(+)-ATPase, H(+)-ATPase, Ca(2+)-ATPase, and carbonic anhydrase activities were assessed in the gills and mantle. Acetylcholinesterase activity was determined in the foot and adductor muscle. The mussels exposed to AgNP accumulated Ag in the gills, hemolymph, and foot, and showed a decrease in hemolymph concentrations of Na(+) and Cl(-), which was associated with the action of Ag ion (Ag(+)). The exposures to TiO(2)NP and FeNP led to the accumulation of Ti and Fe in the hemolymph, respectively, but did not promote additional effects. Accordingly, A. trapesialis showed bioaccumulation potential and susceptibility to AgNP, but was not susceptible to TiO(2)NP and FeNP. Thus, the preferential agricultural use of TiO(2)NP and FeNP over AgNP is highlighted.
ESTHER : Tesser_2022_Chemosphere_309_136664
PubMedSearch : Tesser_2022_Chemosphere_309_136664
PubMedID: 36195123

Title : Effects of copper on an omnivorous (Astyanax altiparanae) and a carnivorous fish (Hoplias malabaricus): A comparative approach - de Paula_2021_Aquat.Toxicol_237_105874
Author(s) : de Paula AA , Risso WE , Martinez C
Ref : Aquat Toxicol , 237 :105874 , 2021
Abstract : Copper is an essential metal for life. However, in excess, it can lead to osmoregulatory disorders and oxidative stress in fish and these effects appear to be species specific. In order to evaluate the effects of copper and to compare the sensitivity of two Neotropical fishes that co-occur in nature as prey (Astyaynax altiparanae) and predator (Hoplias malabaricus), the fish were exposed to three concentrations of Cu (5 microg L(-1), 10 microg L(-1), and 20 microg L(-1)) for 96 h. At the end of the experimental period, copper concentration in tissues, osmoregulatory parameters, oxidative stress biomarkers, plasma glucose, muscle glycogen and acetylcholinesterase activity were evaluated. Fish mortality (25%) was only observed for A. altiparanae exposed to Cu 20 microg L(-1). The results revealed species-specific ionic disturbances. Despite hypocalcemia, H. malabaricus showed an increase in the main gill ATPases, which probably guaranteed the maintenance of plasma Na(+). In A. altiparanae, there was no change in ATPase activity in the gills and hyponatremia was observed at all copper concentrations, as well as a decrease in plasma Cl(-) in the Cu 20 microg L(-1) group. The strategy adopted by H. malabaricus seems to have contributed to the absence of copper accumulation in the tissues, in addition to possibly being related to the absence of oxidative stress in this species. On the other hand, there was an increase in the concentration of copper in the gills, liver, and gastrointestinal tract of A. altiparanae, as well as oxidative stress evidenced by increased lipoperoxidation in the liver and damage to erythrocytes DNA. This work reinforces the idea that copper effects are species specific and that a given concentration may not be safe for different species which can coexist in the same environment.
ESTHER : de Paula_2021_Aquat.Toxicol_237_105874
PubMedSearch : de Paula_2021_Aquat.Toxicol_237_105874
PubMedID: 34090247

Title : Efficient Mimics for Elucidating Zaxinone Biology and Promoting Agricultural Applications - Wang_2020_Mol.Plant_13_1654
Author(s) : Wang JY , Jamil M , Lin PY , Ota T , Fiorilli V , Novero M , Zarban RA , Kountche BA , Takahashi I , Martinez C , Lanfranco L , Bonfante P , de Lera AR , Asami T , Al-Babili S
Ref : Mol Plant , 13 :1654 , 2020
Abstract : Zaxinone is an apocarotenoid regulatory metabolite required for normal rice growth and development. In addition, zaxinone has a large application potential in agriculture, due to its growth-promoting activity and capability to alleviate infestation by the root parasitic plant Striga through decreasing strigolactone (SL) production. However, zaxinone is poorly accessible to the scientific community because of its laborious organic synthesis that impedes its further investigation and utilization. In this study, we developed easy-to-synthesize and highly efficient mimics of zaxinone (MiZax). We performed a structure-activity relationship study using a series of apocarotenoids distinguished from zaxinone by different structural features. Using the obtained results, we designed several phenyl-based compounds synthesized with a high-yield through a simple method. Activity tests showed that MiZax3 and MiZax5 exert zaxinone activity in rescuing root growth of a zaxinone-deficient rice mutant, promoting growth, and reducing SL content in roots and root exudates of wild-type plants. Moreover, these compounds were at least as efficient as zaxinone in suppressing transcript level of SL biosynthesis genes and in alleviating Striga infestation under greenhouse conditions, and did not negatively impact mycorrhization. Taken together, MiZax are a promising tool for elucidating zaxinone biology and investigating rice development, and suitable candidates for combating Striga and increasing crop growth.
ESTHER : Wang_2020_Mol.Plant_13_1654
PubMedSearch : Wang_2020_Mol.Plant_13_1654
PubMedID: 32835886

Title : Single and combined effects of Zn, Mn and Fe on the Neotropical freshwater bivalve Anodontites trapesialis: Bioaccumulation and biochemical biomarkers - Oliveira_2018_Ecotoxicol.Environ.Saf_161_735
Author(s) : Oliveira LF , Cabral MT , Risso WE , Martinez C
Ref : Ecotoxicology & Environmental Safety , 161 :735 , 2018
Abstract : Important concentrations of Zn, Mn and Fe were detected in a stream near a coal mining area and promoted, in field, biomarkers alterations in the bivalve Anodontites trapesialis. In order to understand the isolated and mixed effects of these metals on these Neotropical bivalves, we run short-term experiments under laboratory controlled conditions. After 96h-exposure, tissues (gills, mantle, digestive gland, muscle, hemolymph) were removed for metal bioaccumulation analysis, oxidative stress biomarkers (reactive oxygen species (ROS), total antioxidant capacity, lipoperoxidation (LPO), proteins carbonylation (PC), metallothionein (MT), activity of superoxide dismutase and glutathione S-transferase and hemocytes DNA damage) and cholinesterase (ChE versus ASCh activity) activity evaluation. We run three independent tests. In Zn test, clams were exposed to three concentrations of Zn (0.18mgL(-1), 1.0mgL(-1), 5.0mgL(-1)); in Mn test, clams were exposed to three concentrations of Mn (0.1mgL(-1), 0.5mgL(-1), 5.0mgL(-1)) and in Mix test, clams were exposed to the mixture Zn (1mgL(-1)) +Mn (0.5mgL(-1)), with and without Fe (5.0mgL(-1)). After single exposure to 5.0mgL(-1), Zn bioaccumulated in all tissues, but only in mantle and hemolymph after exposure to 1.0mgL(-1). The increased MT in gills of A. trapesialis exposed to Zn appears to be sufficient to avoid damage, since LPO occurred only in digestive glands from animals exposed to 5.0mgL(-1). We suggested that A. trapesialis had a metabolic suppression in consequence of Mn presence, based on the following results: the decrease of ROS in gills, the decrease of the Zn and Mn concentrations in tissues and the decrease of ChE versus ASCh activity in muscle. Despite this, animals exposed to Mn suffer oxidative damages (LPO and PC) in the mantle and digestive gland and MT increased in the mantle. These results showed A. trapesialis responded differently to each metal and Mn caused more damage. When exposed to Fe, gills level of ROS was increased, despite no changes in metal accumulation occurred. On the other hand, after exposure to the mixtures, tissues bioaccumulated Zn and previously observed damages caused by Mn and Fe disappeared. Consequently, biomarkers were less affected under mixture treatments, demonstrating mixtures effects or responses were not simply a combination of single exposures to Zn, Mn and Fe, but depend on metals toxicokinetics.
ESTHER : Oliveira_2018_Ecotoxicol.Environ.Saf_161_735
PubMedSearch : Oliveira_2018_Ecotoxicol.Environ.Saf_161_735
PubMedID: 29957581

Title : Trends in the prevalence of antipsychotic drug use among patients with Alzheimer's disease and other dementias including those treated with antidementia drugs in the community in the UK: a cohort study - Martinez_2013_BMJ.Open_3_
Author(s) : Martinez C , Jones RW , Rietbrock S
Ref : BMJ Open , 3 : , 2013
Abstract : OBJECTIVE: To investigate the pattern and trends of use of antipsychotics, antidepressants, hypnotics and anxiolytics in Alzheimer's disease and other dementias and in patients treated with antidementia medications. DESIGN: Cohort study with dementia patients formed in the UK Clinical Practice Research Datalink. Participants Patients with incident dementia, between 1995 and 2011 and a reference non-dementia cohort matched on age, gender and date of dementia diagnosis. Two subcohorts included new users of acetylcholinesterase inhibitors (AChEIs) and memantine. The study endpoint was use of antipsychotics, antidepressants, hypnotics and anxiolytics up to 10 years before and 4 years after dementia diagnosis, and for up to 5 years before and 1 year after first use of AChEI or memantine.
RESULTS: 50 349 patients with incident dementia diagnosis and 50 349 matched controls, 10 794 first-time users of AChEI and 669 of memantine. The mean prevalence of antipsychotic use from 1995 to 2011 on diagnosis of dementia was 12.5%, decreasing from 19.9% in 1995 to 7.4% in 2011. There was an increase in antidepressant use (10.7-26.3%) and a small increase in anxiolytic use. The matched cohort showed a lower use of antipsychotics and anxiolytics but a rise in antidepressants (5.9-13.4%). Both groups showed a decrease in hypnotic use. 10.6% of AChEI and 26.3% of memantine users were prescribed antipsychotics, 34.1% and 26.3% antidepressants, 13.2% and 4.1% anxiolytics and 18.4% and 8.3% hypnotics. The slopes for monthly use of antipsychotics were positive in the year leading up to AChEI and memantine use; after treatment initiation the slope for AChEI users continued to increase but at a reduced rate whereas antipsychotic use declined for memantine users.
CONCLUSIONS: The marked reduction in antipsychotic use in dementia is to be welcomed while there was a steady increase in antidepressant use. There was a decline in antipsychotic use after the initiation of memantine.
ESTHER : Martinez_2013_BMJ.Open_3_
PubMedSearch : Martinez_2013_BMJ.Open_3_
PubMedID: 23299113

Title : Pancreatic lipases and their complexes with colipases and inhibitors: crystallization and crystal packing -
Author(s) : Cambillau C , Bourne Y , Egloff MP , Martinez C , van Tilbeurgh H
Ref : Methods Enzymol , 284 :107 , 1997
PubMedID: 9379929

Title : Acyl glycerol hydrolases: inhibitors, interface and catalysis - Cambillau_1996_Curr.Opin.Struct.Biol_6_449
Author(s) : Cambillau C , Longhi S , Nicolas A , Martinez C
Ref : Current Opinion in Structural Biology , 6 :449 , 1996
Abstract : The last five years have witnessed the solution of a large number of lipase structures, which has led, among other insights, to the structural interpretation of the interfacial activation phenomenon in terms of 'lid' opening. This interpretation has been extended this year to include phospholipase A2. Recent structural studies on lipases have provided data on the detailed mechanisms underlying the behaviour of lipases: how they bind to inhibitors or substrates, and what interactions occur between their hydrophobic face and hydrophobic molecules, for example. In addition, studies on cutinase point mutants have shed some light on the role of the oxyanion hole in lipolytic catalysis.
ESTHER : Cambillau_1996_Curr.Opin.Struct.Biol_6_449
PubMedSearch : Cambillau_1996_Curr.Opin.Struct.Biol_6_449
PubMedID: 8794161

Title : Dynamics of Fusarium solani cutinase investigated through structural comparison among different crystal forms of its variants - Longhi_1996_Proteins_26_442
Author(s) : Longhi S , Nicolas A , Creveld L , Egmond M , Verrips CT , de Vlieg J , Martinez C , Cambillau C
Ref : Proteins , 26 :442 , 1996
Abstract : In characterizing mutants and covalently inhibited complexes of Fusarium solani cutinase, which is a 197-residue lipolytic enzyme, 34 variant structures, crystallizing in 8 different crystal forms, have been determined, mostly at high resolution. Taking advantage of this considerable body of information, a structural comparative analysis was carried out to investigate the dynamics of cutinase. Surface loops were identified as the major flexible protein regions, particularly those forming the active-site groove, whereas the elements constituting the protein scaffold were found to retain the same conformation in all the cutinase variants studied. Flexibility turned out to be correlated with thermal motion. With a given crystal packing environment, a high flexibility turned out to be correlated with a low involvement in crystal packing contacts. The high degree of crystal polymorphism, which allowed different conformations with similar energy to be detected, made it possible to identify motions which would have remained unidentified if only a single crystal form had been available. Fairly good agreement was found to exist between the data obtained from the structural comparison and those from a molecular dynamics (MD) simulation carried out on the native enzyme. The crystallographic approach used in this study turned out to be a suitable tool for investigating cutinase dynamics. Because of the availability of a set of closely related proteins in different crystal environments, the intrinsic drawback of a crystallographic approach was bypassed. By combining several static pictures, the dynamics of the protein could be monitored much more realistically than what can be achieved on the basis of static pictures alone.
ESTHER : Longhi_1996_Proteins_26_442
PubMedSearch : Longhi_1996_Proteins_26_442
PubMedID: 8990497
Gene_locus related to this paper: fusso-cutas

Title : Contribution of cutinase serine 42 side chain to the stabilization of the oxyanion transition state - Nicolas_1996_Biochemistry_35_398
Author(s) : Nicolas A , Egmond M , Verrips CT , de Vlieg J , Longhi S , Cambillau C , Martinez C
Ref : Biochemistry , 35 :398 , 1996
Abstract : Cutinase from the fungus Fusarium solani pisi is a lipolytic enzyme able to hydrolyze both aggregated and soluble substrates. It therefore provides a powerful tool for probing the mechanisms underlying lipid hydrolysis. Lipolytic enzymes have a catalytic machinery similar to those present in serine proteinases. It is characterized by the triad Ser, His, and Asp (Glu) residues, by an oxyanion binding site that stabilizes the transition state via hydrogen bonds with two main chain amide groups, and possibly by other determinants. It has been suggested on the basis of a covalently bond inhibitor that the cutinase oxyanion hole may consist not only of two main chain amide groups but also of the Ser42 O gamma side chain. Among the esterases and the serine and the cysteine proteases, only Streptomyces scabies esterase, subtilisin, and papain, respectively, have a side chain residue which is involved in the oxyanion hole formation. The position of the cutinase Ser42 side chain is structurally conserved in Rhizomucor miehei lipase with Ser82 O gamma, in Rhizopus delemar lipase with Thr83 O gamma 1, and in Candida antartica B lipase with Thr40 O gamma 1. To evaluate the increase in the tetrahedral intermediate stability provided by Ser42 O gamma, we mutated Ser42 into Ala. Furthermore, since the proper orientation of Ser42 O gamma is directed by Asn84, we mutated Asn84 into Ala, Leu, Asp, and Trp, respectively, to investigate the contribution of this indirect interaction to the stabilization of the oxyanion hole. The S42A mutation resulted in a drastic decrease in the activity (450-fold) without significantly perturbing the three-dimensional structure. The N84A and N84L mutations had milder kinetic effects and did not disrupt the structure of the active site, whereas the N84W and N84D mutations abolished the enzymatic activity due to drastic steric and electrostatic effects, respectively.
ESTHER : Nicolas_1996_Biochemistry_35_398
PubMedSearch : Nicolas_1996_Biochemistry_35_398
PubMedID: 8555209
Gene_locus related to this paper: fusso-cutas

Title : Horse pancreatic lipase. The crystal structure refined at 2.3 A resolution - Bourne_1994_J.Mol.Biol_238_709
Author(s) : Bourne Y , Martinez C , Kerfelec B , Lombardo D , Chapus C , Cambillau C
Ref : Journal of Molecular Biology , 238 :709 , 1994
Abstract : Pancreatic lipase (EC plays a key role in dietary fat digestion by converting triacylglycerols into 2-monoacylglycerols and free fatty acids in the intestine. Although the crystallographic structures of the human pancreatic lipase and of a human lipase-porcine colipase complex have been solved, no refined structure of pancreatic lipase has yet been published. The crystal structure of the horse enzyme was solved by the molecular replacement method from the model of the human pancreatic lipase and subsequently refined to 2.3 A resolution. The final model contains two molecules of 449 amino acid residues each in the asymmetric unit, 705 well-defined water molecules and two calcium ions. The two molecules in the asymmetric unit of the orthorhombic crystals are related by a 2-fold non-crystallographic symmetry axis as in the case of the human lipase. However, the association between the two molecules in their respective crystal forms is different. The overall molecular structure of the horse lipase is very similar to that of the human enzyme. The horse lipase is made up of two well-defined domains. The N-terminal domain which bears the active centre has a typical alpha/beta hydrolase fold topology. The C-terminal domain which is devoted to colipase binding has a beta-sheet sandwich topology. Comparison of equivalent C alpha atom positions between the final model of the horse lipase and the human lipase structure shows only slight differences which are mainly located in the C-terminal domain. The horse enzyme possesses the common features of the known mammalian and microbial lipases, in particular the "flap" covering the catalytic triad. In addition to more precise information concerning these features, the elucidation of the horse lipase crystal structure allowed us to better understand the structural basis of the kinetic behaviour of pancreatic lipases towards a soluble substrate, p-nitrophenyl acetate, and the different sensitivity of these enzymes towards limited proteolysis.
ESTHER : Bourne_1994_J.Mol.Biol_238_709
PubMedSearch : Bourne_1994_J.Mol.Biol_238_709
PubMedID: 8182745
Gene_locus related to this paper: horse-1plip

Title : Cutinase, a lipolytic enzyme with a preformed oxyanion hole - Martinez_1994_Biochemistry_33_83
Author(s) : Martinez C , Nicolas A , van Tilbeurgh H , Egloff MP , Cudrey C , Verger R , Cambillau C
Ref : Biochemistry , 33 :83 , 1994
Abstract : Cutinases, a group of cutin degrading enzymes with molecular masses of around 22-25 kDa (Kolattukudy, 1984), are also able to efficiently hydrolyse triglycerides (De Geus et al., 1989; Lauwereys et al., 1991), but without exhibiting the interfacial activation phenomenom (Sarda et al., 1958). They belong to a class of proteins with a common structural framework, called the alpha/beta hydrolase fold (Martinez et al., 1992; Ollis et al., 1992). We describe herein the structure of cutinase covalently inhibited by diethyl-p-nitrophenyl phosphate (E600) and refined at 1.9-A resolution. Contrary to what has previously been reported with lipases (Brzozowski et al., 1991; Derewenda et al., 1992; Van Tilbeurgh et al., 1993), no significant structural rearrangement was observed here in cutinase upon the inhibitor binding. Moreover, the structure of the active site machinery, consisting of a catalytic triad (S120, H188, D175) and an oxyanion hole (Q121 and S42), was found to be identical to that of the native enzyme, whereas the oxyanion hole of Rhizomucor lipase (Brzozowski et al., 1991; Derewenda et al., 1992), like that of pancreatic lipase (van Tilbeurgh et al., 1993), is formed only upon enzyme-ligand complex formation. The fact that cutinase does not display interfacial activation cannot therefore only be due to the absence of a lid but might also be attributable to the presence of a preformed oxyanion hole.
ESTHER : Martinez_1994_Biochemistry_33_83
PubMedSearch : Martinez_1994_Biochemistry_33_83
PubMedID: 8286366
Gene_locus related to this paper: fusso-cutas , orysa-LPL1

Title : Engineering cysteine mutants to obtain crystallographic phases with a cutinase from Fusarium solani pisi - Martinez_1993_Protein.Eng_6_157
Author(s) : Martinez C , de Geus P , Stanssens P , Lauwereys M , Cambillau C
Ref : Protein Engineering , 6 :157 , 1993
Abstract : Cutinases are extracellular enzymes involved in the disruption of cutine, an insoluble polyester which covers the surface of plants. They belong to a class of serine esterases that are able to hydrolyse fatty acid esters and emulsified triglycerides as efficiently as lipases, but without displaying interfacial activation. Classical crystallographic methods for obtaining heavy-atom derivatives failed, so the cutinase structure has been solved exclusively by the multiple isomorphous replacement method using four Hg derivatives obtained from mutants S4C, S92C, S120C and S129C. Two of these derivatives behaved as expected: (i) the cys mutant of the catalytic Ser S120C, located at the surface of the active site pocket, leads to a good derivative; and (ii) the Hg atom of the derivative obtained with the S92C mutant is completely accessible to the solvent and occupies two alternative positions--consequently a poor derivative results. In contrast, two mutants show an unexpected behaviour: (i) the Hg atom in the S129C mutant was completely buried 10 A below the protein surface and yielded the best derivative; and (ii) a poor quality derivative was obtained with the S4C mutant. Cys 4 belongs to the disordered propeptide 1-16. The Cys 4 bound Hg atom is located in front of the Asp58 side chain, but neither Cys4 nor parts of the propeptide are clearly visible in the electron density maps of the derivative structure.
ESTHER : Martinez_1993_Protein.Eng_6_157
PubMedSearch : Martinez_1993_Protein.Eng_6_157
PubMedID: 8475042
Gene_locus related to this paper: fusso-cutas

Title : Interfacial activation of the lipase-procolipase complex by mixed micelles revealed by X-ray crystallography - van Tilbeurgh_1993_Nature_362_814
Author(s) : van Tilbeurgh H , Egloff MP , Martinez C , Rugani N , Verger R , Cambillau C
Ref : Nature , 362 :814 , 1993
Abstract : The three-dimensional structure of the lipase-procolipase complex, co-crystallized with mixed micelles of phosphatidylcholine and bile salt, has been determined at 3 A resolution by X-ray crystallography. The lid, a surface helix covering the catalytic triad of lipase, adopts a totally different conformation which allows phospholipid to bind to the enzyme's active site. The open lid is an essential component of the active site and interacts with procolipase. Together they form the lipid-water interface binding site. This reorganization of the lid structure provokes a second drastic conformational change in an active site loop, which in its turn creates the oxyanion hole (induced fit).
ESTHER : van Tilbeurgh_1993_Nature_362_814
PubMedSearch : van Tilbeurgh_1993_Nature_362_814
PubMedID: 8479519
Gene_locus related to this paper: human-PNLIP

Title : Fusarium solani cutinase is a lipolytic enzyme with a catalytic serine accessible to solvent - Martinez_1992_Nature_356_615
Author(s) : Martinez C , de Geus P , Lauwereys M , Matthyssens G , Cambillau C
Ref : Nature , 356 :615 , 1992
Abstract : Lipases belong to a class of esterases whose activity on triglycerides is greatly enhanced at lipid-water interfaces. This phenomenon, called interfacial activation, has a structural explanation: a hydrophobic lid, which at rest covers the catalytic site, is displaced on substrate or inhibitor binding and probably interacts with the lipid matrix. Fusarium solani pisi cutinase belongs to a group of homologous enzymes of relative molecular mass 22-25K (ref. 7) capable of degrading cutin, the insoluble lipid-polyester matrix covering the surface of plants, and hydrolysing triglycerides. Cutinases differ from classical lipases in that they do not exhibit interfacial activation; they are active on soluble as well as on emulsified triglycerides. Cutinases therefore establish a bridge between esterases and lipases. We report here the three-dimensional structure of a recombinant cutinase from F. solani pisi, expressed in Escherichia coli. Cutinase is an alpha-beta protein; the active site is composed of the triad Ser 120, His 188 and Asp 175. Unlike other lipases, the catalytic serine is not buried under surface loops, but is accessible to solvent. This could explain why cutinase does not display interfacial activation.
ESTHER : Martinez_1992_Nature_356_615
PubMedSearch : Martinez_1992_Nature_356_615
PubMedID: 1560844
Gene_locus related to this paper: fusso-cutas , orysa-LPL1

Title : Crystallization and preliminary X-ray study of a recombinant cutinase from Fusarium solani pisi - Abergel_1990_J.Mol.Biol_215_215
Author(s) : Abergel C , Martinez C , Fontecilla-Camps JC , Cambillau C , de Geus P , Lauwereys M
Ref : Journal of Molecular Biology , 215 :215 , 1990
Abstract : Recombinant cutinase from Fusarium solani pisi is expressed and excreted with very high yields in Escherichia coli cultures. Cutinase was crystallized at 20 degrees C using the vapour diffusion technique, with polyethylene glycol 6000 as precipitant. Best crystals were obtained at pH 7.0 with polyethylene glycol 6000 as precipitant. Best crystals were obtained at pH 7.0 with polyethylene glycol at 15 to 20%. They are monoclinic, with space group P2(1) and cell dimensions a = 35.1 A, b = 67.4 A, c = 37.05 A and beta = 94.0 degrees; they diffract beyond 1.5 A resolution. The asymmetric unit contains one molecule of 22,000 Da (Vm = 1.98 A3/Da; 38% water).
ESTHER : Abergel_1990_J.Mol.Biol_215_215
PubMedSearch : Abergel_1990_J.Mol.Biol_215_215
PubMedID: 2213880