Primary structure of H2-proteinase, a non-hemorrhagic metalloproteinase, isolated from the venom of the habu snake, Trimeresurus flavoviridis.
The complete amino acid sequence of and the locations of disulfide bridges in H2-proteinase, a major non-hemorrhagic proteinase isolated from the venom of the habu Trimeresurus flavoviridis, have been determined and compared with those of HR2a, one of the hemorrhagic metalloproteinases in this venom. The strategy involved consisted of structural analysis of peptides in digests with cyanogen bromide, lysyl endopeptidase, trypsin, Staphylococcus aureus V8 protease and thermolysin. Peptides were purified by gel filtration followed by reversed-phase HPLC. H2-proteinase is a non-glycosylated single chain polypeptide consisting of 201 amino acids with an amino-terminal pyroglutamic acid, a calculated molecular weight of 22,991 and a net charge of +14 at neutral pH. There was no evidence of heterogeneity of the sequence. H2-proteinase has a typical zinc-chelating sequence and its overall sequence identity with HR2a is 73.6%. The 3 disulfide bridges in H2-proteinase link Cys-117 to Cys-196, Cys-158 to Cys-180, and Cys-160 to Cys-163, in the same manner as in the case of HR2a. In striking contrast to HR2a, it contains en extra free cysteine residue at position 94 which becomes reactive to a sulfhydryl reagent in the presence of a denaturant.
Department of Biology, Faculty of Science, Kyushu University, Fukuoka.
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Takeya
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Arakawa
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Miyata
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Iwanaga
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Omori-Satoh
D000595:Amino Acid Sequence; D000818:Animals; D003435:Crotalid Venoms / Q000032:analysis*; D004220:Disulfides / Q000032:analysis; D008666:Metalloendopeptidases / Q000032:analysis*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012697:Serine Endopeptidases / Q000378:metabolism; D013329:Structure-Activity Relationship; D013438:Sulfhydryl Compounds / Q000032:analysis
D003435:Crotalid Venoms; D004220:Disulfides; D013438:Sulfhydryl Compounds; C052577:hemorrhagic toxin HR-2a; D012697:Serine Endopeptidases; C036813:lysyl endopeptidase; C061088:H(2)-proteinase; D008666:Metalloendopeptidases