Rescue of ligand binding of a mutant IGF-I receptor by complementation

被引:10
作者
Chakravarty, A
Hinrichsen, J
Whittaker, L
Whittaker, J [1 ]
机构
[1] Case Western Reserve Univ, Dept Nutr, Cleveland, OH 44106 USA
[2] Novo Nordisk AS, Hagedorn Res Inst, Receptor Biol Lab, DK-2820 Gentofte, Denmark
[3] Case Western Reserve Univ, Dept Biochem, Cleveland, OH 44106 USA
关键词
IGF-I; mutant; hybrid receptor; complementation; affinity; dissociation; negative cooperativity;
D O I
10.1016/j.bbrc.2005.03.122
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The IGF-I receptor binds IGF-I with complex kinetics characterized by a curvilinear Scatchard plot, suggesting receptor heterogeneity and apparent negative cooperativity. To explore the molecular mechanisms underlying these properties, we have characterized the binding of a hybrid receptor formed from a wild-type receptor monomer and a mutant receptor monomer devoid of binding activity. Receptor hybrids were generated by transient co-transfection of cDNAs encoding wild-type and Mutant receptors with unique epitope tags. Hybrid receptors were purified front transfected cells by sequential immuno-affinity chromatography and their ligand-binding properties were determined. Complementation produced it hybrid with near wild-type affinity. Dissociation studies demonstrated that the hybrid did not exhibit negative cooperativity. (c) 2005 Elsevier Inc. All rights reserved.
引用
收藏
页码:74 / 77
页数:4
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