Carrick, F.Forbes, B.Wallace, J.2007-05-112007-05-112001Journal of Biological Chemistry, 2001; 276(29):27120-271280021-9258http://hdl.handle.net/2440/28076Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.In the absence of a complete tertiary structure to define the molecular basis of the high affinity binding interaction between insulin-like growth factors (IGFs) and IGF-binding proteins (IGFBPs), we have investigated binding of IGFs by discrete amino-terminal domains (amino acid residues 1-93, 1-104, 1-132, and 1-185) and carboxyl-terminal domains (amino acid residues 96-279, 136-279, and 182-284) of bovine IGFBP-2 (bIGFBP-2). Both halves of bIGFBP-2 bound IGF-I and IGF-II in BIAcore studies, albeit with different affinities (1-132IGFBP-2, KD = 36.3 and 51.8 nM; 136-279IGFBP-2HIS, KD = 23.8 and 16.3 nM, respectively). The amino-terminal half appears to contain components responsible for fast association. In contrast, IGF binding by the carboxyl-terminal fragment results in a more stable complex as reflected by its KD. Furthermore, des(1-3)IGF-I and des(1-6)IGF-II exhibited reduced binding affinity to 1-279IGFBP-2HIS, 1-132IGFBP-2, and 136-279IGFBP-2HIS biosensor surfaces compared with wild-type IGF. A charge reversal at positions 3 and 6 of IGF-I and IGF-II, respectively, affects binding interactions with the amino-terminal fragment and full-length bIGFBP-2 but not the carboxyl-terminal fragment.enJournal article002001065410.1074/jbc.M10131720061886