Multivalent cations and ligand affinity of the type 1 insulin-like growth factor receptor on P2A2-LISN muscle cells.
Mouse P2A2-LISN myoblasts are transfected cells that overexpress the human type 1 insulin-like growth factor (IGF) receptor. Because the type 1 IGF receptor is the major binding site for both IGF-I and IGF-II, this cell line is an excellent model to determine the effect of multivalent cations on ligand binding specifically to this type of receptor. Competitive binding assays were performed to characterize IGF binding and Scatchard analysis to quantify affinity (Ka). 125I-IGF-I, 125I-IGF-II, and 125I-R3-IGF-I bind only to the type 1 IGF receptor on these cells. Zn2+ increased binding of the three ligands to the type 1 IGF receptor by 17 to 35%. Cd2+ significantly increased binding of 125I-IGF-I, although by only 8%. La3+ and Cr3+ did not effect binding. Au3+ decreased IGF binding by approximately 56%. Scatchard analysis produced nonlinear concave-down plots yielding binding constants for high and low affinity sites. Zn2+ increased the strength of only the high affinity sites. Au3+ decreased the affinity of both high and low affinity sites. Zn2+ increased binding with a half-maximal effect between 40 microM and 60 microM. Half-maximal dose of Au3+ was >130 microM. Zinc, gold, and cadmium bind to similar regions within proteins (a zinc-binding motif) and only these cations were found to affect receptor binding indicating similar mechanisms of action. Thus, multivalent cations may alter IGF binding to cell surface receptors ultimately controlling growth. Physiologically this may be especially important for the growth promoting effects of Zn2+.
McCusker
R H
RH
Department of Animal Sciences, The University of Illinois, Urbana, USA. rmccuske@staff.uiuc.edu
Kaleko
M
M
Sackett
R L
RL
D000818:Animals; D001667:Binding, Competitive / Q000502:physiology; D002104:Cadmium / Q000494:pharmacology; D002412:Cations / Q000494:pharmacology; D002478:Cells, Cultured; D002857:Chromium / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006046:Gold / Q000494:pharmacology; D007334:Insulin-Like Growth Factor I / Q000031:analogs & derivatives / Q000494:pharmacology; D007335:Insulin-Like Growth Factor II / Q000494:pharmacology; D007457:Iodine Radioisotopes; D007811:Lanthanum / Q000494:pharmacology; D008024:Ligands; D051379:Mice; D018485:Muscle Fibers, Skeletal / Q000737:chemistry* / Q000166:cytology; D018482:Muscle, Skeletal / Q000166:cytology; D011869:Radioligand Assay; D017526:Receptor, IGF Type 1 / Q000819:agonists / Q000378:metabolism*; D015032:Zinc / Q000494:pharmacology*
D002412:Cations; D007457:Iodine Radioisotopes; D008024:Ligands; C085700:long R(3)-insulin-like growth factor-I; D002104:Cadmium; D002857:Chromium; D007334:Insulin-Like Growth Factor I; D007335:Insulin-Like Growth Factor II; D007811:Lanthanum; D006046:Gold; D017526:Receptor, IGF Type 1; D015032:Zinc