title: Conformation-specific crosslinking of mitochondrial complex I.
abstract: Complex I is the only component of the eukaryotic respiratory chain of which no high-resolution structure is yet available. A notable feature of mitochondrial complex I is the so-called active/de-active conformational transition of the idle enzyme from the active (A) to the de-active, (D) form. Using an amine- and sulfhydryl-reactive crosslinker of 6.8Å length (SPDP) we found that in the D-form of complex I the ND3 subunit crosslinked to the 39 kDa (NDUFA9) subunit. These proteins could not be crosslinked in the A-form. Most likely, both subunits are closely located in the critical junction region connecting the peripheral hydrophilic domain to the membrane arm of the enzyme where the entrance path for substrate ubiquinone is and where energy transduction takes place.
authors: Queen's University Belfast, School of Biological Sciences, Medical Biology Centre, Belfast, UK.
authors: Margherita
authors: M
authors: Ciano
authors: Matthew
authors: M
authors: Fuszard
authors: Heinrich
authors: H
authors: Heide
authors: Catherine H
authors: CH
authors: Botting
authors: Alexander
authors: A
authors: Galkin
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D003432:Cross-Linking Reagents / Q000737:chemistry* / Q000378:metabolism; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D004579:Electron Transport; D042967:Electron Transport Complex I / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D057927:Hydrophobic and Hydrophilic Interactions; D013058:Mass Spectrometry; D024101:Mitochondrial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D013388:Succinimides / Q000737:chemistry / Q000378:metabolism; D014451:Ubiquinone / Q000737:chemistry / Q000378:metabolism
chemical_list: D003432:Cross-Linking Reagents; D024101:Mitochondrial Proteins; D021122:Protein Subunits; D013388:Succinimides; D014451:Ubiquinone; C018151:N-succinimidyl 3-(2-pyridyldithio)propionate; D042967:Electron Transport Complex I; D003545:Cysteine