Document Type
Article
Journal/Book Title
Bio-protocol
Publication Date
11-5-2018
Award Number
NIH 1R15AI124108-01A1
Funder
NIH
Volume
8
Issue
21
First Page
1
Last Page
12
Abstract
ATPases represent a diverse class of enzymes that utilize ATP hydrolysis to support critical biological functions such as driving ion pumps, providing mechanical work, unfolding/folding proteins, and supporting otherwise thermodynamically unfavorable chemical reactions. We have recently shown that the Shigella protein Spa47 is an ATPase that supports protein secretion through its specialized type three secretion apparatus (T3SA), supporting infection of human host cells. Characterizing ATPases, such as Spa47, requires a means to accurately determine enzyme activity (ATP hydrolysis) as a function of time, reaction conditions, and potential cofactors, regulators, inhibitors, etc. Here, we describe a detailed protocol for characterizing the enzyme kinetics of Spa47 using a direct α−32P ATPase assay.
Recommended Citation
Case, Heather B. and Dickenson, Nicholas E., "Kinetic Characterization of the Shigella Type Three Secretion System ATPase Spa47 Using α-32P ATP" (2018). Chemistry and Biochemistry Faculty Publications. Paper 803.
https://digitalcommons.usu.edu/chem_facpub/803
