Factor X-activating glycoprotein of Russell's viper venom. Polypeptide composition and characterization of the carbohydrate moieties.
There is contradictory information regarding the molecular weight and polypeptide chain composition of RVV-X, a glycoprotein in Russell's viper venom that is capable of activating factor X to Xa. We show that RVV-X is a 92,880-Da glycoprotein. It consists of three disulfide-linked polypeptide chains, one heavy chain (alpha-chain, M(r) 57,600) and two light chains (beta- and gamma-chains, M(r) 19,400 and 16,400, respectively). The two light chains seen on SDS-polyacrylamide gel electrophoresis under reducing conditions are two distinct components of the molecule, rather than a heterogeneous mixture of a single chain as previously reported (Takeya, H., Nishida, S., Miyata, T., Kawada, S., Saisaka, Y., Morita, T., and Iwanaga, S. (1992) J. Biol. Chem. 267, 14109-14117). The following evidence supports this conclusion. (i) The two light chains of RVV-X are present in equal proportion. (ii) The estimated molecular weight of an alpha 1 beta 1 gamma 1-structure closely matches the molecular weight determined by matrix-assisted laser desorption mass spectrometry. (iii) The amino acid compositions and NH2-terminal sequences of the beta- and gamma-chains are different. (iv) Although both the beta- and gamma-chains contain one N-linked oligosaccharide chain each, they are glycosylated differentially. RVV-X contains six N-linked oligosaccharides, four in the alpha-chain and one in each of the beta- and gamma-chains. The carbohydrate structures are different from those known for other snake venom glycoproteins, and they resemble closely those in various mammalian glycoproteins. The majority of the oligosaccharides are complex bi-, tri-, and tetraantennary structures, with a small proportion of multiantennary and high mannose-type structures. Two notable structural features of RVV-X oligosaccharides are as follows. (i) Sialic acid residues are linked to beta-galactosyl residues solely by alpha 2,3-linkages, and (ii) bisecting N-acetylglucosamine residues are present in the majority of the oligosaccharides.
Gowda
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Department of Biochemistry and Molecular Biology, Georgetown University Medical Center, Washington, D.C. 20007.
Jackson
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Hensley
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Davidson
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D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002240:Carbohydrate Sequence; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D005170:Factor X / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D006031:Glycosylation; D008567:Membranes, Artificial; D008666:Metalloendopeptidases; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009844:Oligosaccharides / Q000737:chemistry; D010455:Peptides / Q000032:analysis; D011145:Polyvinyls; D017840:Daboia; D014757:Viper Venoms / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D005465:Fluorocarbon Polymers
D010450:Endopeptidases; D005170:Factor X; D006023:Glycoproteins; D008567:Membranes, Artificial; D008666:Metalloendopeptidases; D009844:Oligosaccharides; D010455:Peptides; D011145:Polyvinyls; D014757:Viper Venoms; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers; C033858:russellysin