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The Na+ -translocating NADH:quinone oxidoreductase (Na+ -NQR) from Vibrio cholerae enhances insertion of FeS in overproduced NqrF subunit


Tao, M; Fritz, G; Steuber, J (2008). The Na+ -translocating NADH:quinone oxidoreductase (Na+ -NQR) from Vibrio cholerae enhances insertion of FeS in overproduced NqrF subunit. J. Inorg. Biochem., 102(5-6):1366-1372.

Abstract

The Na+ -translocating NADH:quinone oxidoreductase (Na+ -NQR) from Vibrio cholerae is a membrane-bound, respiratory Na+ pump. Its NqrF subunit contains one FAD and a [2Fe-2S] cluster and catalyzes the initial oxidation of NADH. A soluble variant of NqrF lacking its hydrophobic, N-terminal helix (NqrF´) was produced in V. cholerae wild type and nqr deletion strain. Under identical conditions of growth and induction, the yield of NqrF´ increased by 30% in the presence of the Na+ -NQR. FAD-containing NqrF´ species with or without the FeS cluster were observed, indicating that assembly of the FeS center, but not insertion of the flavin cofactor, was limited during overproduction in V. cholerae. A comparison of these distinct NqrF´ species with regard to specific NADH dehydrogenase activity, pH dependence of activity and thermal inactivation showed that NqrF´ lacking the [2Fe-2S] cluster was less stable, partially unfolded, and therefore prone to proteolytic degradation in V. cholerae. We conclude that the overall yield of NqrF´ critically depends on the amount of fully assembled, FeS-containing NqrF´ in the V. cholerae host cells. The Na+ -NQR is proposed to increase the stability of NqrF´ by stimulating the maturation of FeS centers.

The Na+ -translocating NADH:quinone oxidoreductase (Na+ -NQR) from Vibrio cholerae is a membrane-bound, respiratory Na+ pump. Its NqrF subunit contains one FAD and a [2Fe-2S] cluster and catalyzes the initial oxidation of NADH. A soluble variant of NqrF lacking its hydrophobic, N-terminal helix (NqrF´) was produced in V. cholerae wild type and nqr deletion strain. Under identical conditions of growth and induction, the yield of NqrF´ increased by 30% in the presence of the Na+ -NQR. FAD-containing NqrF´ species with or without the FeS cluster were observed, indicating that assembly of the FeS center, but not insertion of the flavin cofactor, was limited during overproduction in V. cholerae. A comparison of these distinct NqrF´ species with regard to specific NADH dehydrogenase activity, pH dependence of activity and thermal inactivation showed that NqrF´ lacking the [2Fe-2S] cluster was less stable, partially unfolded, and therefore prone to proteolytic degradation in V. cholerae. We conclude that the overall yield of NqrF´ critically depends on the amount of fully assembled, FeS-containing NqrF´ in the V. cholerae host cells. The Na+ -NQR is proposed to increase the stability of NqrF´ by stimulating the maturation of FeS centers.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:04 Faculty of Medicine > Department of Biochemistry
07 Faculty of Science > Department of Biochemistry
Dewey Decimal Classification:570 Life sciences; biology
Language:English
Date:16 January 2008
Deposited On:08 Apr 2008 08:35
Last Modified:05 Apr 2016 12:22
Publisher:Elsevier
ISSN:0162-0134
Publisher DOI:10.1016/j.jinorgbio.2008.01.009
PubMed ID:18289689
Permanent URL: http://doi.org/10.5167/uzh-2318

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