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. Fields of science natural sciencesbiological sciencesmicrobiologybacteriologynatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsnatural sciencesphysical sciencesopticsmicroscopyelectron microscopy Programme(s) FP3-BIOTECH 1 - Specific research and technological development programme (EEC) in the field of biotechnology, 1990-1994 Topic(s) 1.1 - Protein structure and function Call for proposal Data not available Funding Scheme CSC - Cost-sharing contracts Coordinator MRC Laboratory of Molecular Biology Address Hills road CB2 2QH Cambridge United Kingdom See on map EU contribution € 0,00 Participants (3) Sort alphabetically Sort by EU Contribution Expand all Collapse all Commissariat à l'Energie Atomique (CEA) France EU contribution € 0,00 Address Centre d'etudes de grenoble 38041 Grenoble See on map Universidad del País Vasco Spain EU contribution € 0,00 Address S/n,b sarriena s/n 48940 Lejona See on map Universiteit van Amsterdam Netherlands EU contribution € 0,00 Address 127,nieuwe achtergracht 127 1018 WS Amsterdam See on map
