Enzymatic removal of asparagine-linked carbohydrate chains from heterodimer human chorionic gonadotrophin and effect on bioactivity

Craig A H Richard, Mitchell D Creinin, Carolyn J. Kubik, Julie A. DeLoia

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

The native form of human chorionic gonadotropin (hCG) is a heterodimer protein with two asparagine (Asn)-linked carbohydrate chains on each subunit. Removal of the Asn-linked carbohydrate chains from hCG has resulted in hCG variants with consistent antagonistic properties on isolated murine cells. Specific and direct enzymatic removal of these carbohydrate chains from native hCG with resultant antagonistic properties has not been reported. An antagonist to the hCG/luteinising hormone (LH) receptor could be used as an anticancer therapy, emergency contraceptive or for therapeutic resolution of ectopic pregnancies. Therefore, our aim was to use enzymes to specifically remove Asn-linked carbohydrate chains from hCG in the heterodimer form and analyse the resultant bioactivity. Native hCG was treated with endoglycosidases, carbohydrate removal was analysed with electrophoresis and the hCG variants were tested for altered bioactivity with human and murine cells. Endoglycosidases were able to cleave most of the Asn-linked carbohydrate chains from the native hCG. The deglycosylated hCG demonstrated a 75% reduction in bioactivity on a murine Leydig cell line and a 65% reduction in bioactivity on human granulosa cells. These results exemplify a simple and efficient method for creating deglycosylated hCG and provide the most direct evidence for the importance of Asn-linked carbohydrate chains in maintaining hCG bioactivity.

Original languageEnglish (US)
Pages (from-to)933-946
Number of pages14
JournalReproduction, Fertility and Development
Volume19
Issue number8
DOIs
StatePublished - 2007
Externally publishedYes

Keywords

  • Deglycosylation
  • Glycosylation
  • MA-10 cells

ASJC Scopus subject areas

  • Animal Science and Zoology
  • Reproductive Medicine
  • Developmental Biology
  • Endocrinology

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