Effects of the deletion of the Escherichia coli frataxin homologue CyaY on the respiratory NADH:ubiquinone oxidoreductaseReport as inadecuate




Effects of the deletion of the Escherichia coli frataxin homologue CyaY on the respiratory NADH:ubiquinone oxidoreductase - Download this document for free, or read online. Document in PDF available to download.

BMC Biochemistry

, 8:13

First Online: 24 July 2007Received: 15 November 2006Accepted: 24 July 2007

Abstract

BackgroundFrataxin is discussed as involved in the biogenesis of iron-sulfur clusters. Recently it was discovered that a frataxin homologue is a structural component of the respiratory NADH:ubiquinone oxidoreductase complex I in Thermus thermophilus. It was not clear whether frataxin is in general a component of complex I from bacteria. The Escherichia coli homologue of frataxin is coined CyaY.

ResultsWe report that complex I is completely assembled to a stable and active enzyme complex equipped with all known iron-sulfur clusters in a cyaY mutant of E. coli. However, the amount of complex I is reduced by one third compared to the parental strain. Western blot analysis and live cell imaging of CyaY engineered with a GFP demonstrated that CyaY is located in the cytoplasm and not attached to the membrane as to be expected if it were a component of complex I.

ConclusionCyaY plays a non-essential role in the assembly of complex I in E. coli. It is not a structural component but may transiently interact with the complex.

AbbreviationsThe abbreviations used areComplex I, proton-pumping NADH:ubiquinone oxidoreductase

d-NADHdeamino-NADH

DDMn-Dodecyl-β-D-maltopyranoside

decyl-ubiquinone2,3-dimethoxy-5-methyl-6-decyl-benzoquinone

EPRelectron paramagnetic resonance

FMNflavin mononucleotide

Fe-Siron-sulfur

MES2-N-morpholino-ethanesulfonic acid

Tristris-hydroxymethylaminomethane

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2091-8-13 contains supplementary material, which is available to authorized users.

Thomas Pohl, Julia Walter contributed equally to this work.

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Author: Thomas Pohl - Julia Walter - Stefan Stolpe - Joel H Defeu Soufo - Peter L Grauman - Thorsten Friedrich

Source: https://link.springer.com/article/10.1186/1471-2091-8-13







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