Hydrogen exchange (HX) studies have provided critical insight into our understanding of protein folding, structure, and dynamics. More recently, hydrogen exchange mass spectrometry (HX-MS) has become a widely applicable tool for HX studies. The interpretation of the wealth of data generated by HX-MS experiments as well as other HX methods would greatly benefit from the availability of exchange predictions derived from structures or models for comparison with experiment. Most reported computational HX modeling studies have employed solvent-accessible-surface-area based metrics in attempts to interpret HX data on the basis of structures or models. In this study, a computational HX-MS prediction method based on classification of the amide hydrogen bonding modes mimicking the local unfolding model is demonstrated. Analysis of the NH bonding configurations from molecular dynamics (MD) simulation snapshots is used to determine partitioning over bonded and nonbonded NH states and is directly mapped into a protection factor (PF) using a logistics growth function. Predicted PFs are then used for calculating deuteration values of peptides and compared with experimental data. Hydrogen exchange MS data for fatty acid synthase thioesterase (FAS-TE) collected for a range of pHs and temperatures was used for detailed evaluation of the approach. High correlation between prediction and experiment for observable fragment peptides is observed in the FAS-TE and additional benchmarking systems that included various apo/holo proteins for which literature data were available. In addition, it is shown that HX modeling can improve experimental resolution through decomposition of in-exchange curves into rate classes, which correlate with prediction from MD. Successful rate class decompositions provide further evidence that the presented approach captures the underlying physical processes correctly at the single residue level. This assessment is further strengthened in a comparison of residue resolved protection factor predictions for staphylococcal nuclease with NMR data, which was also used to compare prediction performance with other algorithms described in the literature. The demonstrated transferable and scalable MD based HX prediction approach adds significantly to the available tools for HX-MS data interpretation based on available structures and models.
Title: Factors affecting inter-individual variation in human plasma cholinesterase activity: body weight, height, sex, genetic polymorphism and age Brock A, Brock V Ref: Archives of Environmental Contamination & Toxicology, 24:93, 1993 : PubMed
The influence of body weight, height, age and sex on plasma cholinesterase activity (ChE) in 650 males and 437 females with ChE-1 phenotype U (genotype ChEuChEu or ChEuChEs) or UA (genotype ChEuChEa) was studied in a multiple regression model. ChE was not influenced by age (p > 0.01), but, like other liver synthesized plasma enzymes, highly (p < 0.001) influenced by body weight and height. In a logarithmic scale ChE followed a linear model (R = 0.535, p < 0.001) with randomly distributed residuals, InChE = 3.286-0.308 x ChE-1 phenotype-0.104 x sex + 0.00765 x weight - 0.00723 x height (U = 1, UA = 2; male = 1, female = 2; kg; cm). A simplified model based on body-mass index (BMI = weight divided by squared height, kg/m2), InCHE = 2.016-0.308 x ChE-1 phenotype - 0.091 x sex + 0.0230 x BMI, showed the same goodness-of-fit (R = 0.533). In a non-logarithmic scale both multiple regression models failed to fit cases with high ChE activity. A model for a 'standardized' plasma ChE in which the effects of ChE-1 phenotype, sex, body weight and height are eliminated, is proposed to compare ChE in unmatched population groups when using this enzyme activity as a biomarker in environmental or occupational medicine.
Title: Chronic subclinical intake of dietary anticholinesterase agents during the spraying season Lander F, Brock A, Pike E, Hinke K Ref: Food & Chemical Toxicology, 30:37, 1992 : PubMed
The dietary intake of anticholinesterase (anti-ChE) agents was estimated in 331 schoolteachers during the spraying season. Summer plasma-cholinesterase (ChE) activity was compared with the baseline value obtained during winter. Intraindividual plasma-ChE activity varied independently of factors such as drugs, non-malignant diseases, alcohol and smoking. A depressed mean plasma-ChE, indicating an intake of anti-ChE agents (P = 0.04), was observed in individuals who consumed exclusively agriculturally-grown fruits and vegetables without an additional intake of unsprayed, home-grown products. It remains to be determined whether a subclinical but chronic intake of anti-ChE agents in the diet can be hazardous to humans.
Title: Anti-cholinesterase agents uptake during cultivation of greenhouse flowers Lander F, Pike E, Hinke K, Brock A, Nielsen JB Ref: Archives of Environmental Contamination & Toxicology, 22:159, 1992 : PubMed
The cholinesterase (ChE) activities were measured in-season and out-season in a total of 204 greenhouse workers and 360 non-exposed controls. No seasonal ChE variation were observed in the controls, whereas an in-season depression was seen in the workers, indicating an uptake of anti-cholinesterase agents during cultivation of greenhouse flowers in the intervals between sprayings (p = 0.0001). The anti-ChE agents applied seem to persist in the greenhouses and cause continued subtoxic uptake for weeks since last application. Wearing of protective gloves did not prevent the uptake. Thus, chronic percutaneous and oral uptake occurs as a result of cultivation of greenhouse flowers.
Title: Poster: Normal ChE-variation: effects by intra- and inter-individual variance components Brock V, Brock A Ref: In: Cholinesterases: Structure, Function, Mechanism, Genetics, and Cell Biology, (Massoulie J, Barnard EA, Chatonnet A, Bacou F, Doctor BP, Quinn DM) American Chemical Society, Washington, DC:204, 1991 : PubMed
Title: Immunoreactive plasma cholinesterase (EC 3.1.1.8) substance concentration, compared with cholinesterase activity concentration and albumin: inter- and intra-individual variations in a healthy population group Brock A Ref: J Clinical Chemistry Clinical Biochemistry, 28:851, 1990 : PubMed
Substance concentrations of plasma cholinesterase (EC 3.1.1.8) were measured in 94 healthy individuals without occupational exposure to known inhibitors (six samples from each individual). Immunoreactive cholinesterase substance concentrations showed an inter-individual variation corresponding to CVtotal = 22% (mean: 5.01 mg/l, SD: 1.11 mg/l). Intra-individual variations of immunoreactive cholinesterase substance concentration were correlated (r = 0.36) to intra-individual variation of albumin. Estimated by a repeated-measures analysis of variance, the observed intra-individual variation of cholinesterase substance concentration corresponded to CV = 8.8% (SD: 0.44 mg/l), which together with a CVerror = 6% (within and between runs), implies a biological intra-individual variation of cholinesterase substance concentration corresponding to CVintra = 6.4%. Specific catalytic activity (kU/mg immunoreactive cholinesterase) was influenced by the ChE-1 phenotype (phenotype U: 1.58 kU/mg, phenotype UA: 1.22 kU/mg), but not by body weight, height, age, and sex. Observed intra-individual variation of specific catalytic activity corresponded to 6.4% (SD: 0.10 kU/mg), which together with an estimated CVerror = 6.2% implies the biological intra-individual variations of specific catalytic cholinesterase activity to be insignificant. The insignificant CVintra makes specific catalytic cholinesterase activity a rational quantity for evaluation of unexpected fluctuations of cholinesterase activity concentrations.
