Salt stimulation of serum insulin-like growth factor binding protein activity.
Insulin-like growth factors (IGF)-I and -II are bound to carrier or binding proteins in serum. There are at least two classes of binding protein: a high molecular weight complex and a low molecular weight species that is relatively unsaturated. Total binding capacity in serum generally is determined by incubating [125I]IGF with protein that has been stripped of IGF by acid gel filtration. We found that addition of NaCl to the assay increased binding to stripped guinea pig binding protein to about two to four times the level measured in the absence of salt. Stimulation by NaCl was optimal between concentrations of 0.6 and 1.4 M and also was observed when fetal calf or human sera were used as sources of stripped binding protein or when IGF-II was the ligand. Using chloride salts, the order of activity with respect to cations was Na+ greater than K+ greater than Li+. Na2HPO4 at 0.6 M was as stimulatory as 1.2 M NaCl but 0.6 M Na2SO4 was less effective. NH4HCO3 was as effective as NaCl at 0.6 M. Scatchard plots of data from competitive dilution experiments with [125I]IGF-I and unlabeled IGF-I showed that binding was heterogeneous in the absence of 0.6 M NaCl but linear in its presence. NaCl did not stimulate binding when whole serum was used, but after gel filtration of serum on Sephacryl 200 at pH 8, which does not dissociate IGFs from binding protein, binding to individual fractions was stimulated three- to fourfold by NaCl. Fractions stimulated included those containing the large complex or the unsaturated binding protein.(ABSTRACT TRUNCATED AT 250 WORDS)
Palka
J
J
Laboratory of Biochemistry, National Cancer Institute, Bethesda, Maryland 20892.
Peterkofsky
B
B
D000818:Animals; D002352:Carrier Proteins / Q000097:blood* / Q000302:isolation & purification; D002417:Cattle; D002850:Chromatography, Gel / Q000379:methods; D005260:Female; D006168:Guinea Pigs; D018969:Insulin-Like Growth Factor Binding Proteins; D007334:Insulin-Like Growth Factor I / Q000097:blood*; D007335:Insulin-Like Growth Factor II / Q000097:blood*; D007700:Kinetics; D009994:Osmolar Concentration; D011485:Protein Binding; D012965:Sodium Chloride / Q000494:pharmacology; D013002:Somatomedins / Q000097:blood*
D002352:Carrier Proteins; D018969:Insulin-Like Growth Factor Binding Proteins; D013002:Somatomedins; D012965:Sodium Chloride; D007334:Insulin-Like Growth Factor I; D007335:Insulin-Like Growth Factor II