The complete amino acid sequences of two isoproteins of the factor V-activating enzyme (RVV-V) isolated from Vipera russelli (Russell's viper) venom were determined by sequencing S-pyridylethylated derivatives of the proteins and their peptide fragments generated by either chemical (cyanogen bromide and 2-(2-nitrophenylsulfenyl)-3-methyl-3-bromoindolenine) or enzymatic (trypsin, α-chymotrypsin, and lysyl endopeptidase) cleavages. Both enzymes, designated RVV-Vα and RVV-Vγ, consist of 236 amino acid residues and have a N-linked oligosaccharide chain at Asn229. The six amino acid substitutions between RVV-Vα and -Vγ are: Thr22(α)-Ala22(γ), Gly29(α)-Ala29(γ), Gln191(α)-Glu191(γ), Ile192(α)-Met192(γ), Gln193(α)-His193(γ), and Asn224(α)-Ser224(γ). The molecular weights were calculated as 26,182 for RVV-Vα and 26,167 for RVV-Vγ. The sequences of the RVV-V isoproteins exhibited 62% identity with that of batroxobin, a thrombin-like enzyme present in Bothrops atrox venom, and 33% identity with that of human thrombin B chain. The most interesting difference between the structures of RVV-V and other trypsin-type serine proteases is that the conservative Ser214-Trp215-Gly216 sequence (chymotyrpsinogen numbering), considered as the site of antiparallel β-sheet formation between the protein substrate and most serine proteases, has been replaced by the corresponding sequence Ala-Gly-Gly.
|ジャーナル||Journal of Biological Chemistry|
|出版ステータス||出版済み - 1988|
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