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

T. Kuroda, T. Shimamoto, T. Mizushima, T. Tsuchiya

Research output: Contribution to journalArticle

15 Citations (Scopus)

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 K(i) 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.

Original languageEnglish
Pages (from-to)7600-7602
Number of pages3
JournalJournal of Bacteriology
Volume179
Issue number23
Publication statusPublished - 1997
Externally publishedYes

Fingerprint

Vibrio parahaemolyticus
Sodium-Hydrogen Antiporter
Amiloride
Antiporters
Amino Acid Substitution
Site-Directed Mutagenesis
Amino Acid Sequence
Binding Sites

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

Kuroda, T., Shimamoto, T., Mizushima, T., & Tsuchiya, T. (1997). Mutational analysis of amiloride sensitivity of the NhaA Na+/H+ antiporter from Vibrio parahaemolyticus. Journal of Bacteriology, 179(23), 7600-7602.

Mutational analysis of amiloride sensitivity of the NhaA Na+/H+ antiporter from Vibrio parahaemolyticus. / Kuroda, T.; Shimamoto, T.; Mizushima, T.; Tsuchiya, T.

In: Journal of Bacteriology, Vol. 179, No. 23, 1997, p. 7600-7602.

Research output: Contribution to journalArticle

Kuroda, T, Shimamoto, T, Mizushima, T & Tsuchiya, T 1997, 'Mutational analysis of amiloride sensitivity of the NhaA Na+/H+ antiporter from Vibrio parahaemolyticus', Journal of Bacteriology, vol. 179, no. 23, pp. 7600-7602.
Kuroda, T. ; Shimamoto, T. ; Mizushima, T. ; Tsuchiya, T. / Mutational analysis of amiloride sensitivity of the NhaA Na+/H+ antiporter from Vibrio parahaemolyticus. In: Journal of Bacteriology. 1997 ; Vol. 179, No. 23. pp. 7600-7602.
@article{b1b958218501496c815ffc21346a4205,
title = "Mutational analysis of amiloride sensitivity of the NhaA Na+/H+ antiporter from Vibrio parahaemolyticus",
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 K(i) 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.",
author = "T. Kuroda and T. Shimamoto and T. Mizushima and T. Tsuchiya",
year = "1997",
language = "English",
volume = "179",
pages = "7600--7602",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "23",

}

TY - JOUR

T1 - Mutational analysis of amiloride sensitivity of the NhaA Na+/H+ antiporter from Vibrio parahaemolyticus

AU - Kuroda, T.

AU - Shimamoto, T.

AU - Mizushima, T.

AU - Tsuchiya, T.

PY - 1997

Y1 - 1997

N2 - 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 K(i) 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.

AB - 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 K(i) 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.

UR - http://www.scopus.com/inward/record.url?scp=0030696121&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030696121&partnerID=8YFLogxK

M3 - Article

VL - 179

SP - 7600

EP - 7602

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 23

ER -