Skip to main content

Structure and function of NADH : ubiquinone oxidoreductase (complex 1)

Objective

We have undertaken a joint study of the structural features of the mitochondrial, chloroplast and bacterial NADH-CoQ reductase (complex I) from bovine mitochondria, pea chloroplasts and from the bacterium Rhodobacter capsulatus.
Crystallization of a new crystal form of the sub-complex Ilambda fragment of bovine complex I. Sequence analysis of the 42nd subunit of bovine complex I. A 30 Å resolution structure of bovine complex I by single particle analysis electron microscopy. Location of the iron-sulphur clusters 2 in the TYKY subunit of bovine complex I. Demonstration that both a semiquinone radical and a 27structural change in the TYKY subunit of bovine complex I are involved in the coupling of electron transfer to proton translocation. Purification of a protein complex containing the chloroplast encoded protein Ndh K from the thylakoid membranes of chloroplasts from pea plants. Cloning and sequencing of the Rhodobacter nuo operon. Identification of transcriptional signals of Rhodobacter nuo operon. Site directed mutagenesis of Rhodobacter NUO1 (equivalent of bovine TYKY). Isolation of piericidin resistant mutants of Rhodobacter. Partial purification of Rhodobacter complex I.

MAJOR SCIENTIFIC BREAKTHROUGHS:
Structure of bovine complex I at 30 Å resolution. Production of a new 3D-crystal form of the sub-complex 1lambda fragment of bovine complex I. Sequence of a new subunit of bovine complex I. The TYKY subunit of bovine complex I contains Fe-S clusters that are linked via a semiubiquinone radical with the coupling of electron transfer to proton translocation. Purification of a protein complex containing Ndh K from pea chloroplasts. Sequence of the of the Rhodobacter nuo operon. Identification of transcriptional signals of the Rhodobacter nuo operon. Site directed mutants of Rhodobacter NUO1.

Funding Scheme

CSC - Cost-sharing contracts

Coordinator

MRC Laboratory of Molecular Biology
Address
Hills Road
CB2 2QH Cambridge
United Kingdom

Participants (3)

Commissariat à l'Energie Atomique (CEA)
France
Address
Centre D'etudes De Grenoble
38041 Grenoble
Universidad del País Vasco
Spain
Address
S/n,b Sarriena S/n
48940 Lejona
Universiteit van Amsterdam
Netherlands
Address
127,Nieuwe Achtergracht 127
1018 WS Amsterdam