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Sue M. Firth, David R. Clemmons, Robert C. Baxter, Mutagenesis of Basic Amino Acids in the Carboxyl-Terminal Region of Insulin-Like Growth Factor Binding Protein-5 Affects Acid-Labile Subunit Binding, Endocrinology, Volume 142, Issue 5, 1 May 2001, Pages 2147–2150, https://doi.org/10.1210/endo.142.5.8284
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Abstract
Like insulin-like growth factor binding protein-3 (IGFBP-3), IGFBP-5 was recently shown to form ternary complexes with insulin-like growth factor (IGF) and the acid-labile subunit (ALS). Previous studies using IGFBP-5/IGFBP-6 chimeric proteins have identified major and minor ALS binding sites in the carboxyl-terminal and central regions, respectively of IGFBP-5. We now report that ALS binds to IGFBP-3 (Ka = 1.1 ± 0.1 liters/nmol) and IGFBP-5 (Ka = 1.8 ± 0.5 liters/nmol) with similar binding affinities. Using site-specific mutants, we have identified residues K211/R214/K217/R218 within the carboxyl-terminal region of IGFBP-5 as being essential for ALS binding. Mutation of K134R136 or K138K139 in the central region of IGFBP-5 resulted in a small decrease in ALS binding.