Back to Search Start Over

Characterization of inhibitor binding to human kinesin spindle protein by site-directed mutagenesis

Authors :
Carolyn A. Buser
Kelly Hamilton
C. Gary Marshall
Olusegun Williams
Maricel Torrent
Source :
Archives of Biochemistry and Biophysics. 484:1-7
Publication Year :
2009
Publisher :
Elsevier BV, 2009.

Abstract

A number of inhibitors of kinesin spindle protein (KSP) have been described, which are known from X-ray crystallography studies to bind to an induced fit pocket defined by the L5 loop. We describe the characterization of eight mutant forms of KSP in which six residues that line this pocket have been altered. Mutants were analyzed by measuring rates of enzyme catalysis, in the presence and absence of six KSP inhibitors of four diverse structural classes and of varied ATP-competition status. Our analysis was in agreement with the model of binding established by the structural studies and suggests that binding energy is well distributed across functional groups in these molecules. The majority of the mutants retained significant enzymatic activity while diminishing inhibitor binding, indicating potential for the development of drug resistance. These data provide detailed information on interactions between inhibitor and binding pocket at the functional group level and enable the development of novel KSP inhibitors.

Details

ISSN :
00039861
Volume :
484
Database :
OpenAIRE
Journal :
Archives of Biochemistry and Biophysics
Accession number :
edsair.doi.dedup.....58d55d207ef4d4c1fec4acf56381eb84
Full Text :
https://doi.org/10.1016/j.abb.2009.01.015