Structural and membrane binding analysis of the phox homology domain of bem1p - Basis of phosphatidylinositol 4-phosphate specificity

被引:47
作者
Stahelin, Robert V.
Karathanassis, Dimitrios
Murray, Diana
Williams, Roger L.
Cho, Wonhwa
机构
[1] Univ Illinois, Dept Chem, Chicago, IL 60607 USA
[2] Indiana Univ, Sch Med, Dept Biochem & Mol Biol, Bloomington, IN 47405 USA
[3] Univ Notre Dame, Walther Ctr Canc Res, South Bend, IN 46617 USA
[4] Univ Notre Dame, Dept Chem & Biochem, South Bend, IN 46617 USA
[5] MRC, Mol Biol Lab, Cambridge CB2 2QH, England
[6] Cornell Univ, Weill Med Coll, Dept Microbiol & Immunol, New York, NY 10021 USA
基金
英国医学研究理事会;
关键词
D O I
10.1074/jbc.M702861200
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
Phox homology (PX) domains, which have been identified in a variety of proteins involved in cell signaling and membrane trafficking, have been shown to interact with phosphoinositides (PIs) with different affinities and specificities. To elucidate the structural origin of the diverse PI specificity of PX domains, we determined the crystal structure of the PX domain from Bem1p that has been reported to bind phosphatidylinositol 4-phosphate ( PtdIns( 4) P). We also measured the membrane binding properties of the PX domain and its mutants by surface plasmon resonance and monolayer techniques and calculated the electrostatic potentials for the PX domain in the absence and presence of bound PtdIns( 4) P. The Bem1p PX domain contains a signature PI-binding site optimized for PtdIns( 4) P binding and also harbors basic and hydrophobic residues on the membrane-binding surface. The membrane binding of the Bem1p PX domain is initiated by nonspecific electrostatic interactions between the cationicmembrane- binding surface of the domain and anionic membrane surfaces, followed by the membrane penetration of hydrophobic residues. Unlike other PX domains, the Bem1p PX domain has high intrinsic membrane penetrating activity in the absence of PtdIns(4) P, suggesting that the partial membrane penetration may occur before specific PtdIns( 4) P binding and last after the removal of PtdIns( 4) P under certain conditions. This structural and functional study of the PtdIns( 4) P-binding Bem1p PX domain provides new insight into the diverse PI specificities and membrane-binding mechanisms of PX domains.
引用
收藏
页码:25737 / 25747
页数:11
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