Porcine fibrinogen glycopeptides: substrates for detecting endo-beta-N-acetylglucosaminidases F2 and F3(1).
Two different glycopeptides were isolated in high yield from a thermolytic digest of porcine fibrinogen. Edman analysis established their sequences as Val-Glu-Asn(CHO)-Lys and Val-Gly-Glu-Asn(CHO)-Arg. These sequences are nearly identical to the two human fibrinogen glycopeptides, Val-Glu-Asn(CHO)-Lys (gamma-chain), and Met-Gly-Glu-Asn(CHO)-Arg (beta-chain). The predominant carbohydrate moiety of both asialoglycopeptides was a biantennary oligosaccharide with a core alpha(1-->6)-linked fucose as reported earlier (Da Silva et al. (1994) Arch. Biochem. Biophys. 312, 151-157). Both glycopeptides can be dansylated and used as sensitive substrates for Flavobacterium meningosepticum endo-beta-N-acetylglucosaminidases F2 and F3. Porcine fibrinogen represents the best source for substrates with this oligosaccharide type that can be reliably produced in multimicromole quantities.
Plummer
T H
TH
Division of Molecular Medicine, New York State Department of Health, Albany 12201-0509, USA.
Phelan
A W
AW
Tarentino
A L
AL
D000118:Acetylglucosaminidase / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D005340:Fibrinogen / Q000378:metabolism*; D006020:Glycopeptides / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D008969:Molecular Sequence Data; D013379:Substrate Specificity; D013552:Swine
D006020:Glycopeptides; D007527:Isoenzymes; D005340:Fibrinogen; D000118:Acetylglucosaminidase