A complete system for identifying inhibitors of creatine kinase B.
We have developed a complete system for discovery of lead compounds as inhibitors of creatine kinase B. In this article, we describe production and purification of the recombinant protein, conditions and features of an optimized high-throughput screening assay, and results of our implementation of the system using a diverse compound library.
Towler
E M
EM
SAIC Frederick, NCI-FCRDC, Frederick, Maryland, 21702, USA. Eric.Towler.b@Bayer.com
Wilson
L K
LK
Zhou
Y C
YC
Ma
T S
TS
Fisher
R J
RJ
D001483:Base Sequence; D001921:Brain / Q000201:enzymology; D003001:Cloning, Molecular; D003402:Creatine Kinase / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D017931:DNA Primers / Q000235:genetics; D015195:Drug Design; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D066298:In Vitro Techniques; D007527:Isoenzymes; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification
D017931:DNA Primers; D004791:Enzyme Inhibitors; D007527:Isoenzymes; D011994:Recombinant Proteins; D003402:Creatine Kinase