Electrophoretic and isoelectric focusing analysis of human recombinant alpha2-HS glycoprotein produced in insect cells: Analysis of the post-translational events

Laszlo Kalabay, Savita Mathur, Sophie Bobin, Philippe Arnaud

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Alpha2-HS glycoprotein (AHSG) is a human serum glycoprotein Synthesized by liver cells. It is a natural inhibitor of the insulin receptor tyrosine kinase activity. We produced this protein in insect cells by using a recombinant baculovirus expressing the whole coding sequence of the protein. By analyzing AHSG on isoelectric focusing and on sodium dodecyl sulfate (SDS) gels, followed by immunoblot, AHSG produced in insect cells was found to be phosphorylated and to possess the connecting peptide between the A and the B chains. The same features were found in the protein produced by Hep3B, a human liver cell line that synthesizes AHSG. By contrast, no phosphorylation could be detected in AHSG present in normal human plasma, and the connecting peptide was clipped. As the protein produced in insect cells is active on insulin receptors, in contrast to the plasma protein, our results suggest that the biological activity of the protein may be associated with its single chain form together with its phosphorylation.

Original languageEnglish
Pages (from-to)529-532
Number of pages4
JournalELECTROPHORESIS
Volume17
Issue number3
DOIs
Publication statusPublished - Mar 1 1996

Keywords

  • Alpha2-HS glycoprotein
  • Baculovirus expression system
  • Insulin receptor tyrosine kinase
  • Isoelectric focusing
  • Posttranslational modification

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry

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