title: The external calcium-dependent NADPH dehydrogenase from Neurospora crassa mitochondria.
abstract: We have inactivated the nuclear gene coding for a putative NAD(P)H dehydrogenase from the inner membrane of Neurospora crassa mitochondria by repeat-induced point mutations. The respiratory rates of mitochondria from the resulting mutant (nde-1) were measured, using NADH or NADPH as substrates under different assay conditions. The results showed that the mutant lacks an external calcium-dependent NADPH dehydrogenase. The observation of NADH and NADPH oxidation by intact mitochondria from the nde-1 mutant suggests the existence of a second external NAD(P)H dehydrogenase. The topology of the NDE1 protein was further studied by protease accessibility, in vitro import experiments, and in silico analysis of the amino acid sequence. Taken together, it appears that most of the NDE1 protein extends into the intermembrane space in a tightly folded conformation and that it remains anchored to the inner mitochondrial membrane by an N-terminal transmembrane domain.
authors.affiliation: Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto, Portugal.
authors.forename: A M
authors.initials: AM
authors.lastname: Melo
authors.forename: M
authors.initials: M
authors.lastname: Duarte
authors.forename: I M
authors.initials: IM
authors.lastname: Møller
authors.forename: H
authors.initials: H
authors.lastname: Prokisch
authors.forename: P L
authors.initials: PL
authors.lastname: Dolan
authors.forename: L
authors.initials: L
authors.lastname: Pinto
authors.forename: M A
authors.initials: MA
authors.lastname: Nelson
authors.forename: A
authors.initials: A
authors.lastname: Videira
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D008928:Mitochondria / Q000201:enzymology*; D009252:NADPH Dehydrogenase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D009492:Neurospora crassa / Q000201:enzymology*
chemical_list: D000255:Adenosine Triphosphate; D009252:NADPH Dehydrogenase; D002118:Calcium