Insulin and IGF binding by IGFBP-3 fragments derived from proteolysis, baculovirus expression and normal human urine

Peter Vorwerk, Yoshitaka Yamanaka, Anna Spagnoli, Youngman Oh, Ron G. Rosenfeld

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Recombinant human IGFBP-3 was proteolysed with different concentrations of plasmin for various periods of time. The major IGFBP-3 fragment resulting from this digestion migrated at ca. 15 kDa in nonreducing SDS-PAGE. Following the identification of this fragment as an N-terminal IGFBP-3 fragment, by use of N-terminus-specific monoclonal antibody and amino acid sequence analysis, we constructed and expressed a similar fragment in a baculovims expression system. The fragments resulting from plasmin digestion, as well as the baculovirus-expressed recombinant human IGFBP-31-97, retain weak IGF binding and show specific insulin binding on cross-linking and western ligand blot. RhIGFBP-31-97 can inhibit insulin receptor autophosphorylation in insulin receptor-overexpressing NIH 3T3 cells. Insulin and IGF binding to IGFBP-3 fragments could be further demonstrated in normal urine. These data indicate the physiological significance of IGFBP-3 fragments derived from proteolysis in vivo.

Original languageEnglish (US)
Pages (from-to)1392-1395
Number of pages4
JournalJournal of Clinical Endocrinology and Metabolism
Volume83
Issue number4
DOIs
StatePublished - Jan 1 1998

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Proteolysis
Insulin-Like Growth Factor Binding Protein 3
Baculoviridae
Urine
Insulin
Fibrinolysin
Insulin Receptor
Digestion
Insulin-Like Growth Factor Binding Proteins
NIH 3T3 Cells
Protein Sequence Analysis
Polyacrylamide Gel Electrophoresis
Western Blotting
Monoclonal Antibodies
Ligands
Amino Acids

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Biochemistry
  • Endocrinology
  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Insulin and IGF binding by IGFBP-3 fragments derived from proteolysis, baculovirus expression and normal human urine. / Vorwerk, Peter; Yamanaka, Yoshitaka; Spagnoli, Anna; Oh, Youngman; Rosenfeld, Ron G.

In: Journal of Clinical Endocrinology and Metabolism, Vol. 83, No. 4, 01.01.1998, p. 1392-1395.

Research output: Contribution to journalArticle

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