Application of Single-Molecule Spectroscopy in Studying Enzyme Kinetics and Mechanism

Jue Shi, Joseph Dertouzos, Ari Gafni, Duncan Steel

Research output: Chapter in book/report/conference proceedingChapterpeer-review

Abstract

This chapter reviews recent developments in the application of single-molecule spectroscopy (SMS) to studies of enzyme kinetics and mechanism. Protocols for conducting single-molecule experiments on enzymes, based largely on the experience in our laboratory, are provided, including methods of sample preparation, instrumentation, and data analysis. We also address general issues related to the design of meaningful single-molecule experiments and include specific examples of the application of SMS to enzyme studies, which reveal new and intriguing aspects of enzyme behavior, including static and dynamic heterogeneity, as well as subunit cooperativity. Finally, we discuss the advantages of employing single-molecule approach in obtaining new information beyond ensemble studies.
Original languageEnglish
Title of host publicationFluorescence Spectroscopy
EditorsLudwig Brand, Michael L. Johnson
PublisherAcademic Press
Pages129-157
Number of pages29
ISBN (Print)9780123745866
DOIs
Publication statusPublished - 25 Dec 2008

Publication series

NameMethods in Enzymology
Volume450
ISSN (Print)0076-6879

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