The Enzymes and the Enzyme Complexes of the Mitochondrial Oxidative Phosphorylation System

Hatefi, Youssef; Ragan, C I; Galante, Yves M.

doi:10.1007/978-1-4684-4604-3_1

Cited by 33 publications

(13 citation statements)

References 290 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the first studies the complex was desintegrated with chaotropic anions. This fractionation yielded three subcomplexes; the flavoprotein fragment with three subunits, one of them containing FMN and the NADH-binding site; the ironprotein fragment which comprises six subunits, some of them containing iron-sulfur clusters and a hydrophobic fraction, consisting mainly of membrane proteins of unknown function [30,31].…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Immunopurification of a subcomplex of the NAD(P)H‐plastoquinone‐oxidoreductase from the cyanobacterium Synechocystis sp. PCC6803

et al. 1993

View full text Add to dashboard Cite

An antibody against the NDH‐K subunit of the NAD(P)H‐dehydrogenase from the cyanobacterium Synechocystis sp. PCC6803 was used to isolate a subcomplex of the enzyme from Triton X‐100 solubilized total membranes by immunoaffinity chromatography. The isolated subcomplex consisted of seven major polypeptides with molecular masses of 43, 27, 24, 21, 18, 14 and 7 kDa. The amino‐terminal amino acid sequences of the polypeptides were determined. By comparing the sequences with the amino acid sequences deduced from DNA, three proteins were identified as NDH‐H (43 kDa), NDH‐K (27 kDa) and NDH‐I (24 kDa). A fourth subunit (NDH‐J, 21 kDa) was identified by Western blot analysis with an NDH‐J antibody.

show abstract

Section: Discussionmentioning

confidence: 99%

“…The polypeptide composition of this fragment was initially thought to consist of six proteins with molecular masses of 75, 49, 30, 18, 15 and 13 kDa [30,31]. The 49 kDa protein is homologous to NDH-H [32] and the 30 kDa protein to NDH-J [33,34].…”

Section: Discussionmentioning

confidence: 99%

Immunopurification of a subcomplex of the NAD(P)H‐plastoquinone‐oxidoreductase from the cyanobacterium Synechocystis sp. PCC6803

et al. 1993

View full text Add to dashboard Cite

show abstract

“…Enzyme markers for the vacuole membrane (a-mannosidase), the A total of 100 spontaneous sucrose-fermenting mutants were isolated with strains SEY2101, SEY2102, SEY2108, and SEY2109 bearing the pCYI433 plasmid. The selection was performed in the presence of antimycin A, an inhibitor of yeast respiratory metabolism (27). All but 2 of these independently obtained mutants exhibited sucrose-fermenting phenotypes that were recessive in heterozygous diploids.…”

Section: Constructionmentioning

confidence: 99%

Isolation of yeast mutants defective in protein targeting to the vacuole.

Bankaitis¹,

Johnson²,

Emr³

1986

Proc. Natl. Acad. Sci. U.S.A.

364

309

View full text Add to dashboard Cite

We have constructed a PRCI-SUC2 gene fusion that directs the synthesis in Saccharomyces cerevisiae of a hybrid polypeptide consisting of a 433-residue amino-terminal domain derived from the yeast vacuolar protease carboxypeptidase Y (CPY; EC 3.4.16.1) and a 511-residue carboxyl-terminal domain derived from the secreted yeast enzyme invertase (EC 3.2.1.26). Fractionation data indicated that this amount of CPY primary sequence is sufficient to quantitatively divert invertase to the yeast vacuole. The phenotypic consequence of localizing active invertase to the vacuole has enabled us to select for mutants that "mislocalize" the hybrid protein to the cell surface. The corresponding mutations that lead to this effect are all trans-acting and recessive, and they define at least eight complementation groups. These vacuolar protein targeting (vpt) mutants also exhibit hybrid protein independent defects in wild-type CPY delivery to the yeast vacuole. Precursor forms of CPY accumulate in the mutants and are secreted into the yeast periplasm and extracellular medium. The vpt mutants should provide useful information pertaining to the mechanisms by which yeast cells regulate vacuolar protein traffic.

show abstract

“…Data from the bovine (11), the Paracoccus (12), and the E. coli (13) enzymes have indicated that the 51-kDa subunit binds NAD(H) and contains FMN and a tetranuclear cluster (center N3), and the 24-kDa subunit contains a binuclear cluster (likely center N1a). The bovine IP is also water-soluble, and is composed of seven major polypeptides with molecular masses of 75, 49,30,18,15,13, and 11 kDa, of which the last two comigrate at M r ϳ13,000 upon SDS-polyacrylamide gel electrophoresis (14,15). Only the 75-kDa subunit houses iron-sulfur clusters, and data from the Paracoccus enzyme have shown that the bacterial analogue of this subunit contains a tetranuclear, a binuclear (likely center N1b), and possibly another tetranuclear iron-sulfur cluster (16).…”

mentioning

confidence: 99%

Mitochondrial NADH-Ubiquinone Oxidoreductase (Complex I)

Yamaguchi

Belogrudov²,

Hatefi

1998

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

It has been shown that treatment of bovine mitochondrial complex I (NADH-ubiquinone oxidoreductase) with NADH or NADPH, but not with NAD or NADP, increases the susceptibility of a number of subunits to tryptic degradation. This increased susceptibility involved subunits that contain electron carriers, such as FMN and iron-sulfur clusters, as well as subunits that lack electron carriers. Results shown elsewhere on changes in the cross-linking pattern of complex I subunits when the enzyme was pretreated with NADH or NADPH (Belogrudov, G., and Hatefi, Y. (1994) Biochemistry 33, 4571-4576) also indicated that complex I undergoes extensive conformation changes when reduced by substrate. Furthermore, we had previously shown that in submitochondrial particles the affinity of complex I for NAD increases by >20-fold in electron transfer from succinate to NAD when the particles are energized by ATP hydrolysis. Together, these results suggest that energy coupling in complex I may involve protein conformation changes as a key step.In addition, it has been shown here that treatment of complex I with trypsin in the presence of NADPH, but not NADH or NAD(P), produced from the 39-kDa subunit a 33-kDa degradation product that resisted further hydrolysis. Like the 39-kDa subunit, the 33-kDa product bound to a NADP-agarose affinity column, and could be eluted with a buffer containing NADPH. It is possible that together with the acyl carrier protein of complex I the NADP(H)-binding 39-kDa subunit is involved in intramitochondrial fatty acid synthesis.Among the enzyme complexes of the mitochondrial electron transport/oxidative phosphorylation system, complex I (NADH-ubiquinone oxidoreductase) has the most complicated structure and mechanism of action. The enzyme is composed of Ն43 subunits (1, 2), and contains one FMN and 5 EPR-visible iron-sulfur clusters, designated centers N1a, N1b, N2, N3, and N4 (3). Chemical analysis of iron and sulfide suggested the possible presence of 8 iron-sulfur clusters per FMN (4). These results agreed with more recent data on the amino acid sequences of complex I subunits, in which 8 cysteine motifs were found for ligating 6 tetranuclear and 2 binuclear iron-sulfur clusters (1, 2).Early studies showed that bovine complex I could be resolved into three domains, designated as flavoprotein (FP), 1 iron-sulfur protein (IP), and hydrophobic protein (HP) (5, 6). This tridomain architecture appears to be the structural design of complex I from Neurospora crassa mitochondria (Ն35 subunits) (7), Escherichia coli plasma membrane (13 subunits) (8, 9), and possibly Paracoccus denitrificans (14 subunits) (10). Bovine FP is water-soluble; it is composed of three subunits with molecular masses of 51, 24, and 9 kDa, and it contains per mol 1 FMN, a tetranuclear iron-sulfur cluster, and a binuclear iron-sulfur cluster. Data from the bovine (11), the Paracoccus (12), and the E. coli (13) enzymes have indicated that the 51-kDa subunit binds NAD(H) and contains FMN and a tetranuclear cluster (center N3), and the 24-kDa s...

show abstract

The Enzymes and the Enzyme Complexes of the Mitochondrial Oxidative Phosphorylation System

Cited by 33 publications

References 290 publications

Immunopurification of a subcomplex of the NAD(P)H‐plastoquinone‐oxidoreductase from the cyanobacterium Synechocystis sp. PCC6803

Immunopurification of a subcomplex of the NAD(P)H‐plastoquinone‐oxidoreductase from the cyanobacterium Synechocystis sp. PCC6803

Isolation of yeast mutants defective in protein targeting to the vacuole.

Mitochondrial NADH-Ubiquinone Oxidoreductase (Complex I)

Contact Info

Product

Resources

About