Defining balanced conditions for inhibitor screening assays that target bisubstrate enzymes

Journal of Biomolecular Screening

Volumn 14, Issue 2, 2009, Pages 111-120

  Yang, Jingsong ^a   Copeland, Robert A ^a   Lai, Zhihong ^a  

^a GLAXOSMITHKLINE   (United States)

Author keywords

Balanced conditions; Bisubstrate; Enzymatic assay; High throughput screening; IC50; Ki; Optimal screening conditions

Indexed keywords

ENZYME INHIBITOR;

ARTICLE; COMPETITIVE INHIBITION; DRUG MECHANISM; DRUG POTENCY; DRUG SCREENING; DRUG TARGETING; ENZYME KINETICS; ENZYME SUBSTRATE; HIGH THROUGHPUT SCREENING; IC 50; PRACTICE GUIDELINE; PRIORITY JOURNAL; THEORETICAL STUDY;

EID: 61849124218     PISSN: 10870571     EISSN: 1552454X     Source Type: Journal    
DOI: 10.1177/1087057108328763     Document Type: Article

References (14)

1. Hopkins AL, Groom CR: The druggable genome. Nat Rev Drug Discov 2002 ; 1: 727-730.
2. Drews J.: Drug discovery: a historical perspective. Science 2000 ; 287: 1960-1964.
3. Zhang JH, Chung TD, Oldenburg KR: A simple statistical parameter for use in evaluation and validation of high throughput screening assays. J Biomol Screen 1999 ; 4: 67-73.
4. Macarron R., Hertzberg RP: Design and implementation of high throughput screening assays. Methods Mol Biol 2002 ; 190: 1-29.
5. Walters WP, Namchuk M.: Designing screens: how to make your hits a hit. Nat Rev Drug Discov 2003 ; 2: 259-266.
6. Copeland RA: Mechanistic considerations in high-throughput screening. Anal Biochem 2003 ; 320: 1-12.
7. Wu G., Yuan Y., Hodge CN: Determining appropriate substrate conversion for enzymatic assays in high-throughput screening. J Biomol Screen 2003 ; 8: 694-700.
8. Chene P.: Challenges in design of biochemical assays for the identification of small molecules to target multiple conformations of protein kinases. Drug Discov Today 2008 ; 13: 522-529.
9. Segal IH: Enzyme Kinetics: Behavior and Analysis of Rapid Equilibrium and Steady-State Enzyme Systems. New York: John Wiley, 1975.
10. Copeland RA: Evaluation of Enzyme Inhibitors in Drug Discovery: A Guide for Medicinal Chemists and Pharmacologists. New York: John Wiley, 2005.
11. Copeland RA: Enzymes: A Practical Introduction to Structure, Mechanism, and Data Analysis. New York: John Wiley, 2000.
12. Case A., Ni J., Yeh LA, Stein RL: Development of a mechanism-based assay for tissue transglutaminase: results of a high-throughput screen and discovery of inhibitors. Anal Biochem 2005 ; 338: 237-244.
13. Cheng Y., Prusoff WH: Relationship between the inhibition constant (Ki) and the concentration of inhibitor which cause 50 percent inhibition (IC50) of an enzymatic reaction. Biochem Pharmacol 1973 ; 22: 3099-3108.
14. Lueck JD, Ellison WR, Fromm HJ: The point of convergence of sequential double reciprocal plots as a criterion of bireactant enzyme kinetic mechanisms: evaluation of the yeast hexokinase reaction. FEBS Lett 1973 ; 30: 321-324.