title: Properties of glycosylated and non-glycosylated human recombinant IGF binding protein-3 (IGFBP-3).
abstract: The interaction of insulin-like growth factor-I with glycosylated and nonglycosylated human recombinant IGFBP-3 was studied by competition binding and by realtime biospecific interaction analysis (BIA). No significant difference was found in the affinity of the two forms of recombinant IGFBP-3 for IGF-I. Combining the results from the two different methods of binding analysis, a dissociation constant of 50 +/- 14 pM was determined. This value compares favorably to that reported for IGF-I binding to natural human plasma-derived IGFBP-3. Subcutaneous injection into rats of IGF-I either bound to glycosylated or nonglycosylated IGFBP-3 yielded an area under the curve (AUC) that was essentially the same for either form of IGFBP-3 and was twice as large as the AUC obtained after injection of IGF-I in the free form. Circulating IGF-I peak levels were reached in less than 1 h post-injection for free IGF-I, 4 and 8 h post-injection for the nonglycosylated and glycosylated IGF-I/IGFBP-3 complexes respectively. Neutral gel filtration of rat serum samples 4 and 8 h post-injection revealed that the IGF-I delivered bound to the nonglycosylated IGFBP-3 circulated as a 40-50 kDa complex at the 4 h time point and as a 130-140 kD complex at the 8 h time point. IGF-I delivered as a complex with glycosylated IGFBP-3 was found to circulate as a 140 kD complex at 4 and 8 h post-injection.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Celtrix Pharmaceuticals, Inc., Santa Clara, California 95052.
authors.forename: A
authors.initials: A
authors.lastname: Sommer
authors.forename: S K
authors.initials: SK
authors.lastname: Spratt
authors.forename: G P
authors.initials: GP
authors.lastname: Tatsuno
authors.forename: T
authors.initials: T
authors.lastname: Tressel
authors.forename: R
authors.initials: R
authors.lastname: Lee
authors.forename: C A
authors.initials: CA
authors.lastname: Maack
mesh_terms: D000818:Animals; D016466:CHO Cells; D002352:Carrier Proteins / Q000235:genetics / Q000493:pharmacokinetics / Q000502:physiology*; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D006224:Cricetinae; D004926:Escherichia coli; D005947:Glucose / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D007279:Injections, Subcutaneous; D018969:Insulin-Like Growth Factor Binding Proteins; D007334:Insulin-Like Growth Factor I / Q000378:metabolism / Q000493:pharmacokinetics; D008297:Male; D011485:Protein Binding; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000378:metabolism / Q000493:pharmacokinetics; D013002:Somatomedins
chemical_list: D002352:Carrier Proteins; D018969:Insulin-Like Growth Factor Binding Proteins; D011994:Recombinant Proteins; D013002:Somatomedins; D007334:Insulin-Like Growth Factor I; D005947:Glucose