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Glycosylation of the thrombin-like serine protease ancrod fromAgkistrodon rhodostoma venom. Oligosaccharide substitution pattern at eachN-glycosylation site

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Abstract

In a previous study, we determined the structures of the glycans present in ancrod, a thrombin-like serine protease from the venom of the Malayan pit viperAgkistrodon rhodostoma (Pfeifferet al. (1992)Eur J Biochem 205:961–78). In order to allocate the various carbohydrate chains to distinctN-glycosylation sites of the molecule, we have now isolated individual glycopeptides. Peptide moieties were identified after deglycosylation with peptide-N 4-(N-acetyl-β-glucosaminyl)asparagine amidase F by amino acid analysis and sequencing. Liberated oligosaccharides were assigned to the previously deduced carbohydrate structures by high performance liquid chromatography. Although only quantitative differences were observed, the results indicate that each glycosylation site of ancrod carries its characteristic oligosaccharide pattern. Furthermore, all potential sites were shown to be substituted by carbohydrates.

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Abbreviations

HPAE-HPLC:

high pH anion exchange HPLC

RP-HPLC:

reversed phase HPLC

PNGase-F:

peptide-N 4-(N-acetyl-β-glucosaminyl)asparagine amidase F

PAD:

pulsed amperometric detection.

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Correspondence to Rudolf Geyer.

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Pfeiffer, G., Linder, D., Strube, K. et al. Glycosylation of the thrombin-like serine protease ancrod fromAgkistrodon rhodostoma venom. Oligosaccharide substitution pattern at eachN-glycosylation site. Glycoconjugate J 10, 240–246 (1993). https://doi.org/10.1007/BF00702206

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Keywords

  • glycoprotein
  • glycopeptides
  • N-linked oligosaccharides
  • snake venom