Location of disulfide bonds within the sequence of human serum cholinesterase.
Human serum cholinesterase was digested with pepsin under conditions which left disulfide bonds intact. Peptides were isolated by high pressure liquid chromatography, and those containing disulfide bonds were identified by a color assay. Peptides were characterized by amino acid sequencing and composition analysis. Human serum cholinesterase contains 8 half-cystines in each subunit of 574 amino acids. Six of these form three internal disulfide bridges: between Cys65-Cys92, Cys252-Cys263, and Cys400-Cys519. A disulfide bond with Cys65 rather than Cys66 was inferred by homology with Torpedo acetylcholinesterase. Cys571 forms a disulfide bridge with Cys571 of an identical subunit. This interchain disulfide bridge is four amino acids from the carboxyl terminus. A peptide containing the interchain disulfide is readily cleaved from cholinesterase by trypsin (Lockridge, O., and La Du, B. N. (1982) J. Biol. Chem. 257, 12012-12018), suggesting that the carboxyl terminus is near the surface of the globular tetrameric protein. The disulfide bridges in human cholinesterase have exactly the same location as in Torpedo californica acetylcholinesterase. There is one potential free sulfhydryl in human cholinesterase at Cys66, but this sulfhydryl could not be alkylated. Comparison of human cholinesterase, and Torpedo and Drosophila acetylcholinesterases to the serine proteases suggests that the cholinesterases constitute a separate family of serine esterases, distinct from the trypsin family and from subtilisin.
D000110:Acetylcholinesterase / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D002802:Cholinesterases / Q000097:blood* / Q000235:genetics / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D003545:Cysteine; D004220:Disulfides / Q000032:analysis; D004330:Drosophila / Q000201:enzymology; D006801:Humans; D007202:Indicators and Reagents; D008969:Molecular Sequence Data; D010434:Pepsin A; D010446:Peptide Fragments / Q000032:analysis; D012689:Sequence Homology, Nucleic Acid; D014101:Torpedo
D004220:Disulfides; D007202:Indicators and Reagents; D010446:Peptide Fragments; D000110:Acetylcholinesterase; D002802:Cholinesterases; D010434:Pepsin A; D003545:Cysteine