Back to Search
Start Over
Characterization of inhibitor binding to human kinesin spindle protein by site-directed mutagenesis
- 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.
- Subjects :
- chemistry.chemical_classification
Sequence Homology, Amino Acid
Protein Conformation
Molecular Sequence Data
Mutant
Mutagenesis
Allosteric regulation
Biophysics
Kinesins
Crystallography, X-Ray
Binding, Competitive
Biochemistry
Enzyme catalysis
Adenosine Triphosphate
Enzyme
chemistry
Biocatalysis
Mutagenesis, Site-Directed
Humans
Kinesin
Amino Acid Sequence
Human Kinesin Spindle Protein
Site-directed mutagenesis
Molecular Biology
Subjects
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