T. Nikai et al., PRIMARY STRUCTURE OF A COAGULANT ENZYME, BILINEOBIN, FROM AGKISTRODONBILINEATUS VENOM, Archives of biochemistry and biophysics, 318(1), 1995, pp. 89-96
The amino acid sequence and disulfide bridge location of the coagulant
enzyme, named bilineobin, isolated from the venom of Agkistrodon bili
neatus was determined by Edman sequencing of the peptides derived from
digests with cyanogen bromide, clostripain, Staphylococcus aureus V8
protease, trypsin, and chymotrypsin. This enzyme has a molecular weigh
t of 57,000 Da by sodium dodecyl sulfate-polyacrylamide gel electropho
resis; however, bilineobin consists of 235 amino acids and has a calcu
lated molecular weight of 26,481, The enzyme contains fucose, GlcNAc,
galactose, mannose and NeuAc and six N-linked glycosylation consensus
sites. The carboxyterminal amino acid, proline, was determined using c
arboxypeptidase Y. The six disulfide bonds of bilineobin link Cys(78)
to Cys(234), Cys(120) to Cys(188), Cys(178) to Cys(203), Cys(7) to Cys
(141), Cys(152) to Cys(167), and Cys(28) to Cys(44). The amino acid se
quence similarity to flavoxobin (T. C. Shieh et al., 1988, J. Biochem
(Tokyo) 103, 596-605) and batroxobin (N. Itoh ed al., 1987, J. Biol. C
hem. 262, 3132-3135) was 67%, The deglycosylated enzyme more rapidly g
enerated fibrinopeptide A than native bilineobin. (C) 1995 Academic Pr
ess, Inc.