title: Role of the conserved cysteine residues of the 11.5 kDa subunit in complex I catalytic properties.
abstract: Mitochondrial complex I exists as a mixture of two inter-convertible forms: active (A) and de-activated (D), the latter being sensitive to SH-modifying compounds. To investigate if the conserved cysteine-rich 11.5 kDa subunit of Neurospora crassa complex I is involved in this process, we subjected the corresponding genomic DNA to site-directed mutagenesis. The four cysteine residues of the subunit were separately substituted with serine residues and the resulting proteins were independently expressed in a null-mutant strain. All of the obtained mutant strains were able to assemble a complex I with similar kinetic properties to those observed in the wild-type enzyme, indicating that none of the cysteine residues of the 11.5 kDa protein is individually relevant for the A/D transition process. Diminished amounts of assembled complex I seem to be the major effect of these specific mutations. The cysteine residues are likely important to the acquisition and stabilization of the correct 11.5 kDa protein conformation and this is reflected in the assembly/stability of complex I.
authors.affiliation: IBMC Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua do Campo Alegre 823, Porto, Portugal. imarques@ibmc.up.pt
authors.forename: Isabel
authors.initials: I
authors.lastname: Marques
authors.forename: Alexandra V
authors.initials: AV
authors.lastname: Ushakova
authors.forename: Margarida
authors.initials: M
authors.lastname: Duarte
authors.forename: Arnaldo
authors.initials: A
authors.lastname: Videira
mesh_terms: D003545:Cysteine / Q000378:metabolism*; D042967:Electron Transport Complex I / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008928:Mitochondria / Q000737:chemistry / Q000378:metabolism; D009154:Mutation; D009492:Neurospora crassa / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation
chemical_list: D042967:Electron Transport Complex I; D003545:Cysteine