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Publication Abstract from PubMed

IQGAP1 is a multi-domain scaffold protein involved in many cellular processes. We have determined the crystal structure of an N-terminal fragment spanning residues 1-191 (CHDF hereafter) which contains the entire calponin homology domain. The structure of the CHDF is very similar to other Type 3 calponin homology domains like those from calponin, Vav, and the yeast IQGAP1 ortholog Rng2. However, in the crystal two CHDF molecules form a "head-to-head" or parallel dimer through mostly hydrophobic interactions. Binding experiments indicate that the CHDF binds to both F-actin and Ca2+/calmodulin, but binding is mutually exclusive. Based on the structure, two dimer interface substitutions were introduced. While CHDFL157D disrupts the dimer in gel filtration experiments, oxidized CHDFK161C stabilizes the dimer. These results imply that the CHDF forms the same dimer in solution that is seen in the crystal structure. The disulfide-stabilized dimer displays reduced F-actin binding in sedimentation assays, and shows no binding to Ca2+/calmodulin in isothermal titration calorimetry (ITC) experiments indicating that interface residues are utilized for both binding events. The Calmodulin Target Database predicts that residues 93KK94 are important for CaM binding, and indeed the 93EE94 double mutation displays reduced binding to Ca2+/calmodulin in ITC experiments. Our results indicate that the CHDF dimer interface is used for both F-actin and Ca2+/calmodulin binding, and 93KK94, near the interface, are also used for Ca2+/calmodulin binding. These results are also consistent with full-length IQGAP1 forming a parallel homodimer.

The IQGAP1 N-terminus Forms Dimers and the Dimer Interface is Required for Binding F-actin and Ca2+/calmodulin.,Liu J, Kurella VB, Lecour L, Vanagunas T, Worthylake DK Biochemistry. 2016 Oct 31. PMID:27798963^[2]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.