Abstract
The usualE u1 and atypicalE a1 human pseudocholinesterases (acylocholine acylhydrolase, EC 3.1.1.8) were purified to homogeneity. The active-site serine residue was conjugated with diisopropyl fluorophosphate and digested with trypsin. The tryptic peptide containing the active site was isolated by gel filtration followed by two-dimensional paper chromatography and electrophoresis. The amino acid sequence of the active site peptide obtained from the usualE u1 enzyme was found to be Gly-Glu-Ser-Ala-Gly-Ala-Ser-Ala-Val-Ser-Leu. A remarkable structural homology exists between the human and the horse enzymes in their active sites. From the difference in electrophoretic mobility of the active-site peptides obtained from the usual and atypical enzymes, the probable structure of the atypical human enzyme was deduced as Gly-His-Ser-Ala-Gly-Ala-Ser-Ala-Val-Ser-Leu.
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References
Cuatrecasas, P. (1970). Protein purification by affinity chromatography.J. Biol. Chem. 2453059;
Davies, R. O., Marton, A. V., and Kalow, W. (1960). The action of normal and atypical cholinesterase of human serum upon a series of esters of choline.Can. J. Biochem. Physiol. 38545.
Goedde, H.- W., Gehring, D., and Hofmann, R. A. (1965). On the problem of a “silent gene” in pseudocholinesterase polymorphism.Biochim. Biophys. Acta 107391.
Haupt, V. H., Heide, K., Zwisler, O., and Schwick, H. G. (1966). Isolierung und physikalisch-chemische Charakterisierung der Cholinesterase aus human Serum.Blut 1465.
Hodgkin, W. E., Giblett, E. R., Levine, H., Bauer, W., and Motulsky, A. G. (1965). Complete pseudocholinesterase deficiency: Genetic and immunologic characterization.J. Clin. Invest. 44486.
Huang, I- Y., Rubinfien, E., and Yoshida, A. (1980). Complete amino acid sequence of human phosphoglycerate kinase.J. Biol. Chem. 2556408.
Jansz, H. S., Brons, D., and Warringa, M. G. P. J. (1959a). Chemical nature of DFP-binding site of pseudocholinesterase.Biochim. Biophys. Acta 34573.
Jansz, H. S., Posthumus, C. H., and Cohen, J. A. (1959b). On the active site of horse-liver ali esterase. II. Amino acid sequence in the DFP-binding site of the enzyme.Biochim. Biophys. Acta 33396.
Jones, R. T. (1964). Structural studies of aminoethylated hemoglobins by automatic peptide chromatography.Cold Spring Harbor Symp. Quant. Biol. 29297.
Kalow, W., and Davies, R. O. (1958). The activity of various esterase inhibitors towards atypical human serum cholinesterase.Biochem. Pharmacol. 1183.
Kalow, W., and Genest, K. (1957). A method for the detection of atypical forms of human serum cholinesterase. Determination of dibucaine numbers.Can. J. Biochem. Physiol. 35339.
Kalow, W., and Gunn, D. R. (1959). Some statistical data on atypical cholinesterase of human serum.Ann. Hum. Genet. (Lond.) 23239.
Kalow, W., and Lindsay, H. A. (1955). A comparison of optical and manometric methods for assay of human serum cholinesterase.Can. J. Biochem. 33568.
Kalow, W., and Staron, N. (1957). On distribution and inheritance of human serum cholinesterase, as indicated by dibucaine numbers.Can. J. Biochem. 351305.
Lehmann, H., and Ryan, E. (1956). The familial incidence of low pseudocholinesterase level.Lancet 2124.
Lockridge, O., and La Du, B. N. (1978). Comparison of atypical and usual human serum cholinesterase.J. Biol. Chem. 253361.
Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951). Protein measurements with the Folin phenol reagent.J. Biol. Chem. 193265.
Mendez, E., and Lai, C. Y. (1975). Regeneration of amino acid from thiazolines formed in the Edman degradation.Anal. Biochem. 6847.
Muensch, H., Goedde, H.-W., and Yoshida, A. (1976). Human serum cholinesterase subunits and number of active sites of the major component.Eur. J. Biochem. 70217.
Muensch, H., Yoshida, A., Altland, K., Jensen, W., and Goedde, H.- W. (1978). Structural difference at the active site of dibucaine resistant variant of human plasma cholinesterase.Am. J. Hum. Genet. 30302.
Naughton, M. A., Sanger, F., Hartley, B. S., and Shaw, D. W. (1960). Amino acid sequence around the reactive serine residue of some proteolytic enzyme.Biochem. J. 77149.
Schaffer, N. K., May, C. S., and Summerson, W. H. (1954). Serine Phosphoric acid from diisopropylphosphoryl derivative of eel cholinesterase.J. Biol. Chem. 206201.
Schaffer, N. K., Michel, H. O., and Bridges, A. F. (1973). Amino acid sequence in the region of the reactive serine residue of eel acetylcholinesterase.Biochemistry 122946.
Summers, M. R., Smythers, G. W., and Oroszlan, S. (1973). Thin-layer chromatography of sub-nanomole amounts of phenylthiohydantoin (PTH) amino acids on polyamide sheets.Anal. Biochem. 53624.
Thompson, J. C., and Whittaker, M. (1966). A study of the pseudocholinesterase in 78 cases of apnea following suxamethionium.Acta Genet. 16206.
Warren, L. (1959). The thiobarbituric acid assay of sialic acid.J. Biol. Chem. 2341971.
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This work was supported by Grants HL-15125 and HL-29515 from the National Institutes of Health.
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Yamato, K., Huang, IY., Muensch, H. et al. Amino acid sequence of the active site of human pseudocholinesterase. Biochem Genet 21, 135–145 (1983). https://doi.org/10.1007/BF02395397
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DOI: https://doi.org/10.1007/BF02395397