Enzymatic characterization of the enteropathogenic Escherichia coli type III secretion ATPase EscN

Angel Andrade; Juan Pablo Pardo; Norma Espinosa; Gerardo Pérez-Hernández; Bertha González-Pedrajo

doi:10.1016/j.abb.2007.09.020

Enzymatic characterization of the enteropathogenic Escherichia coli type III secretion ATPase EscN

Arch Biochem Biophys. 2007 Dec 1;468(1):121-7. doi: 10.1016/j.abb.2007.09.020. Epub 2007 Sep 29.

Authors

Angel Andrade¹, Juan Pablo Pardo, Norma Espinosa, Gerardo Pérez-Hernández, Bertha González-Pedrajo

Affiliation

¹ Departamento de Genética Molecular, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Ap. Postal 70-243, México, DF 04510, Mexico.

PMID: 17964526
DOI: 10.1016/j.abb.2007.09.020

Abstract

Type III secretion is a transport mechanism by which bacteria secrete proteins across their cell envelope. This protein export pathway is used by two different bacterial nanomachines: the flagellum and the injectisome. An indispensable component of these secretion systems is an ATPase similar to the F1-ATPase beta subunit. Here we characterize EscN, an enteropathogenic Escherichia coli type III ATPase. A recombinant version of EscN, which was fully functional in complementation tests, was purified to homogeneity. Our results demonstrate that EscN is a Mg2+-dependent ATPase (kcat 0.35 s(-1)). We also define optimal conditions for the hydrolysis reaction. EscN displays protein concentration-dependent activity, suggesting that the specific activity changes with the oligomeric state of the protein. The presence of active oligomers was revealed by size exclusion chromatography and native gel electrophoresis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Biological Transport, Active / physiology
Ca(2+) Mg(2+)-ATPase / chemistry*
Ca(2+) Mg(2+)-ATPase / metabolism*
Enzyme Activation
Enzyme Stability
Escherichia coli / enzymology*
Substrate Specificity

Substances

Ca(2+) Mg(2+)-ATPase