Topological study of Vibrio alginolyticus NhaB Na+/H+ antiporter using gene fusions in escherichia coli cells

Hiromi Enomoto; Tsutomu Unemoto; Mitsuaki Nishibuchi; Etana Padan; Tatsunosuke Nakamura

doi:10.1016/S0005-2736(97)00245-9

Topological study of Vibrio alginolyticus NhaB Na⁺/H⁺ antiporter using gene fusions in escherichia coli cells

Hiromi Enomoto, Tsutomu Unemoto, Mitsuaki Nishibuchi, Etana Padan, Tatsunosuke Nakamura^*

^*Corresponding author for this work

Alexander Silberman Institute of Life Sciences

Research output: Contribution to journal › Article › peer-review

20 Scopus citations

Abstract

NhaB, an Na⁺/H⁺ antiporter, of Vibrio alginolyticus is a 528-amino- acid protein. Hydropathy profile-based computer analysis predicted that the NhaB might contain up to 13 membrane-spanning domains. To examine this hypothesis, we applied the phoA fusion method to the cloned nhaB gene. Eighteen plasmid-borne nhaB-phoA fusion genes were constructed in Escherichia coli cells and the alkaline phosphatase activity and expression level of the fusion proteins analyzed. These results and the results obtained with additional constructs indicated that V. alginolyticus NhaB has a unique topology consisting of nine transmembrane segments with the N-terminus in the cytoplasm and the C-terminus in the periplasm.

Original language	English
Pages (from-to)	77-86
Number of pages	10
Journal	Biochimica et Biophysica Acta - Biomembranes
Volume	1370
Issue number	1
DOIs	https://doi.org/10.1016/S0005-2736(97)00245-9
State	Published - 6 Mar 1998

Keywords

(Vibrio alginolyticus)
Marine bacterium
NhaB gene
Sodium ion/proton antiporter
Topology

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10.1016/S0005-2736(97)00245-9

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title = "Topological study of Vibrio alginolyticus NhaB Na+/H+ antiporter using gene fusions in escherichia coli cells",

abstract = "NhaB, an Na+/H+ antiporter, of Vibrio alginolyticus is a 528-amino- acid protein. Hydropathy profile-based computer analysis predicted that the NhaB might contain up to 13 membrane-spanning domains. To examine this hypothesis, we applied the phoA fusion method to the cloned nhaB gene. Eighteen plasmid-borne nhaB-phoA fusion genes were constructed in Escherichia coli cells and the alkaline phosphatase activity and expression level of the fusion proteins analyzed. These results and the results obtained with additional constructs indicated that V. alginolyticus NhaB has a unique topology consisting of nine transmembrane segments with the N-terminus in the cytoplasm and the C-terminus in the periplasm.",

keywords = "(Vibrio alginolyticus), Marine bacterium, NhaB gene, Sodium ion/proton antiporter, Topology",

author = "Hiromi Enomoto and Tsutomu Unemoto and Mitsuaki Nishibuchi and Etana Padan and Tatsunosuke Nakamura",

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AU - Unemoto, Tsutomu

AU - Nishibuchi, Mitsuaki

AU - Padan, Etana

AU - Nakamura, Tatsunosuke

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KW - (Vibrio alginolyticus)

KW - Marine bacterium

KW - NhaB gene

KW - Sodium ion/proton antiporter

KW - Topology

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Topological study of Vibrio alginolyticus NhaB Na⁺/H⁺ antiporter using gene fusions in escherichia coli cells

Abstract

Keywords

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