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J Clin Endocrinol Metab. 1998 Apr;83(4):1392-5.

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

Author information

1
Dept. Pediatrics NRC5, Oregon Health Sciences University, Portland 97201-3042, USA.

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 baculovirus expression system. The fragments resulting from plasmin digestion, as well as the baculovirus-expressed recombinant human IGFBP-3(1-97), retain weak IGF binding and show specific insulin binding on cross-linking and western ligand blot. RhIGFBP-3(1-97) 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.

PMID:
9543173
DOI:
10.1210/jcem.83.4.4858
[Indexed for MEDLINE]

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