A method for [3H]mannose labeling of Asn-linked oligosaccharides on recombinant glycoproteins synthesized in Xenopus oocytes

Alan B. Cantor, Stuart Kornfeld

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

We have developed an efficient method for labeling the Asn-linked oligosaccharides of recombinant glycoproteins synthesized in Xenopus laevis oocytes. By coinjecting GDP-[3,4-3H]mannose with mRNA for human cathepsin D, it was possible to incorporate as much as 1800 cpm per oocyte into each of the two Asn-linked oligosaccharides of this glycoprotein. Overall, about 50% of the microinjected GDP-[3,4-3H]mannose was incorporated into Asn-linked oligosaccharides, a 10-fold greater value than that obtained when [2-3H]mannose was microinjected. Less than 10% of the injected GDP-[3,4-3H]mannose was metabolized to water or converted to amino acids. This technique should facilitate studies of Asn-linked oligosaccharide biosynthesis, processing, and structure in recombinant proteins synthesized in Xenopus oocytes.

Original languageEnglish
Pages (from-to)220-226
Number of pages7
JournalAnalytical Biochemistry
Volume205
Issue number2
DOIs
StatePublished - Sep 1992

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