ERIC Number: EJ1166952
Record Type: Journal
Publication Date: 2018-Jan
Abstractor: As Provided
Bisubstrate Kinetics of Glutathione S-Transferase: A Colorimetric Experiment for the Introductory Biochemistry Laboratory
Stefanidis, Lazaros; Scinto, Krystal V.; Strada, Monica I.; Alper, Benjamin J.
Journal of Chemical Education, v95 n1 p146-151 Jan 2018
Most biochemical transformations involve more than one substrate. Bisubstrate enzymes catalyze multiple chemical reactions in living systems and include members of the transferase, oxidoreductase, and ligase enzyme classes. Working knowledge of bisubstrate enzyme kinetic models is thus of clear importance to the practicing biochemist. However, such models are infrequently explored in the undergraduate biochemistry laboratory. This deficiency suggests the need for well-defined, tractable, and economical methods for characterization of bisubstrate enzyme activity. The application of an established real-time colorimetric assay to investigate kinetic parameters of glutathione transfer by glutathione S-transferase is described. The reported method relies on commercially available materials and uses experimental conditions that are suited to application in the introductory biochemistry laboratory. Investigation of bisubstrate kinetic parameters was performed as part of a three-week laboratory experiment that emphasized protein extraction, purification, and characterization, leading to guided student development of bisubstrate enzyme kinetic models.
Descriptors: Introductory Courses, Kinetics, Biochemistry, Models, Undergraduate Students, Science Laboratories, Laboratory Experiments
Division of Chemical Education, Inc and ACS Publications Division of the American Chemical Society. 1155 Sixteenth Street NW, Washington, DC 20036. Tel: 800-227-5558; Tel: 202-872-4600; e-mail: firstname.lastname@example.org; Web site: http://pubs.acs.org/jchemeduc
Publication Type: Journal Articles; Reports - Descriptive
Education Level: Higher Education
Authoring Institution: N/A
Grant or Contract Numbers: N/A