O-Linked L-Fucose Is Present in Desmodus rotundus Salivary Plasminogen Activator
| Autor: | Peter Bringmann, Werner Reutter, Rolf Nuck, Detlef Grunow, Christoph Kannicht, Günther Baude, Martin Gohlke, Peter Donner |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
Threonine
Glycosylation Molecular Sequence Data CHO Cells Transfection Peptide Mapping Biochemistry Gas Chromatography-Mass Spectrometry Mass Spectrometry Fucose Plasminogen Activators chemistry.chemical_compound Chiroptera Cricetinae Animals Humans Amino Acid Sequence Saliva Molecular Biology Peptide sequence Chromatography High Pressure Liquid chemistry.chemical_classification Sequence Homology Amino Acid biology Cell Biology biology.organism_classification Molecular biology Recombinant Proteins Glycopeptide Amino acid chemistry Tissue Plasminogen Activator Desmodus rotundus Cattle Plasminogen activator |
| Zdroj: | Journal of Biological Chemistry. 271:7381-7386 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.271.13.7381 |
| Popis: | DSPAalpha1 (Desmodus rotundus salivary plasminogen activator), a plasminogen activator from the saliva of the vampire bat Desmodus rotundus, is an effective thrombolytic agent. An unusual type of posttranslational modification, in which L-fucose is O-glycosidically linked to threonine 61 in the epidermal growth factor domain was found for natural DSPAalpha1 and its recombinant form isolated from Chinese hamster ovary cells. In the present study a combination of carbohydrate and amino acid composition analysis, amino acid sequencing, and mass spectrometry revealed that the L-fucose is bound to residues 56-68 of DSPAalpha1. The amino acid sequence of this glycosylation site agreed with the suggested consensus sequence Cys-Xaa-Xaa-Gly-Gly-Ser/Thr-Cys described for other proteins. Anew strategy for the identification of the modified amino acid was established. Direct evidence for the occurrence of fucosyl-threonine was obtained by mass spectrometry after digestion of the glycopeptide with a mixture of peptidases. On the basis of these results, DSPAalpha1 is a suitable model for studying the influence of O-fucosylation on clearance rates, particularly in comparative studies with the identically fucosylated and structurally related tissue plasminogen activator. |
| Databáze: | OpenAIRE |
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