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PMID: 9393731 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Mutational analysis of amiloride sensitivity of the NhaA Na+/H+ antiporter from Vibrio parahaemolyticus.

Journal of bacteriology ·Vol. 179 ·No. 23 ·1997-12-00 ·Pages 7600-2

Kuroda T, Shimamoto T, Mizushima T, Tsuchiya T

Abstract

The activity of the NhaA Na+/H+ antiporter of Vibrio parahaemolyticus is inhibited by amiloride. We found an amino acid sequence in the NhaA that was identical to a putative amiloride binding domain of the Na+/H+ exchanger in mammalian cells. We constructed mutant NhaAs that had amino acid substitutions in the putative amiloride binding domain by site-directed mutagenesis. These include V62L (Val62 replaced by Leu), F63Y, F64Y, and L65F. Most mutant NhaAs showed decreased sensitivity for amiloride. Among these, the F64Y mutant NhaA showed the least amiloride sensitivity, with a Ki value 7 to 10 times greater than that in the wild type. Thus, the sequence between residues V62 and L65 in NhaA, especially F64, is very important for the inhibitory effect of amiloride on the antiporter.

MeSH Terms
Amiloride/pharmacology Binding Sites DNA Mutational Analysis Escherichia coli Proteins Lithium/metabolism Protons Sodium/metabolism Sodium-Hydrogen Exchangers/antagonists & inhibitors,genetics Vibrio parahaemolyticus
Chemicals
Escherichia coli Proteins NhaA protein, E coli Protons Sodium-Hydrogen Exchangers Amiloride Lithium Sodium
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Kuroda T
Department of Microbiology, Faculty of Pharmaceutical Sciences, Okayama University, Tsushima, Japan.
Shimamoto T
Mizushima T
Tsuchiya T
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.179.23.7600-7602.1997
Published
1997-12-00
Pages
7600-2
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC179717
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.179.23.7600-7602.1997 PMC Full Text View journal details
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