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Interdependence of two NarK domains in a fused nitrate/nitrite transporter

Goddard, Alan D. and Moir, James W. B. and Richarson, David J. and Ferguson, Stuart J. (2008) Interdependence of two NarK domains in a fused nitrate/nitrite transporter. Molecular Microbiology, 70 (3). pp. 667-681. ISSN 0950-382X

Full content URL: http://dx.doi.org/10.1111/j.1365-2958.2008.06436.x

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Item Type:Article
Item Status:Live Archive

Abstract

Nitrate uptake is essential for various bacterial processes and combines with nitrite export to form the usual initial steps of denitrification, a process that reduces nitrate to dinitrogen gas. Although many bacterial species contain NarK-like transporters that are proposed to function as either nitrate/proton symporters or nitrate/nitrite antiporters based on sequence homology, these transporters remain, in general, poorly characterized. Several bacteria appear to contain a transporter that is a fusion of two NarK-like proteins, although the significance of this arrangement remains elusive. We demonstrate that NarK from Paracoccus denitrificans is expressed as a fusion of two NarK-like transporters. NarK1 and NarK2 are separately capable of supporting anaerobic denitrifying growth but with growth defects that are partially mitigated by coexpression of the two domains. NarK1 appears to be a nitrate/proton symporter with high affinity for nitrate and NarK2 a nitrate/nitrite antiporter with lower affinity for nitrate. Each transporter requires two conserved arginine residues for activity. A transporter consisting of inactivated NarK1 fused to active NarK2 has a dramatically increased affinity for nitrate compared with NarK2 alone, implying a functional interaction between the two domains. A potential model for nitrate and nitrite transport in P. denitrificans is proposed.

Keywords:nitrate transporter, Paracoccus denitrificans
Subjects:C Biological Sciences > C700 Molecular Biology, Biophysics and Biochemistry
Divisions:College of Science > School of Life Sciences
ID Code:6830
Deposited On:15 Nov 2012 21:21

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