Ligands to the 2Fe iron‐sulfur center in succinate dehydrogenase

Aevarsson, Arnthór; Hederstedt, Lars

doi:10.1016/0014-5793(88)80757-9

Cited by 13 publications

(5 citation statements)

References 37 publications

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“…Electron paramagnetic resonance (EPR) analysis of deletion mutants of the Bacillus subtilis complex II suggest that the IP N-terminal end interacts with the FP protein (Hederstedt etal., 1985;Aevarsson & Hederstedt, 1988). This conclusion is also inferred from bioenergetic considerations which suggest that the [2Fe-2S] center, located in the N-terminal end, may be the immediate acceptor of electrons from the reduced flavin moiety (Cammack, 1986).…”

Section: Discussionmentioning

confidence: 99%

Studies on the assembly of complex II in yeast mitochondria using chimeric human/yeast genes for the iron-sulfur protein subunit

Saghbini¹,

Pl²,

Ie³

1994

Biochemistry

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A series of chimeric human/yeast IP genes were constructed in order to investigate domains of the iron-sulfur protein (IP) that are important for assembly and/or activity of complex II of the electron transport system in Saccharomyces cerevisiae. These genes were expressed in a respiration-deficient yeast mutant in which the endogenous IP gene had been disrupted. Substitutions at the N-terminus were tolerable. Substituting the region covering the first iron-sulfur center [2Fe-2S] had no effect on assembly, while activity decreased 2-5-fold. The addition of seven amino acids from the human peptide, including four charged residues, at the C-terminus did not perturb either assembly or activity. A region between the first and second cysteine clusters was identified which when substituted caused a complete failure in the assembly of complex II. It includes a 15 amino acid stretch which shows the greatest variability between species. Larger substitutions including this segment failed as well. Exchanging the region between the second and third cysteine clusters making up the [4Fe-4S] and [3Fe-4S] centers enabled transformants to grow on nonfermentable carbon sources, yet no SDH activity was observed in vitro. The IP and FP proteins accumulate to wild-type levels in these mutants. We speculate that the lack of observed activity is due to the lability of iron-sulfur centers in isolated, broken mitochondria.

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Section: Discussionmentioning

confidence: 99%

Studies on the assembly of complex II in yeast mitochondria using chimeric human/yeast genes for the iron-sulfur protein subunit

Saghbini¹,

Pl²,

Ie³

1994

Biochemistry

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“…These different types of iron-sulfur centers are displayed in the amino acid sequence of the iron-sulfur protein containing 11 conserved cysteine residues (10 in Escherichia coli SdhB) which are supposed to function as ligands of the metal clusters. Four cysteine residues (three in E. coli) are arranged in a cluster close to the N terminus and ligand center 1, as deduced from studies of mutant enzymes with truncated iron-sulfur subunits (3,27,36), as well as from site-directed mutagenesis (62). The tetranuclear center 2 and trinuclear center 3 are supposed to be ligated by cysteine residues arranged in two clusters near the C terminus.…”

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confidence: 99%

A succinate dehydrogenase with novel structure and properties from the hyperthermophilic archaeon Sulfolobus acidocaldarius: genetic and biophysical characterization

1997

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The sdh operon of Sulfolobus acidocaldarius DSM 639 is composed of four genes coding for the 63.1-kDa flavoprotein (SdhA), the 36.5-kDa iron-sulfur protein (SdhB), and the 32.1-kDa SdhC and 14.1-kDa SdhD subunits. The four structural genes of the sdhABCD operon are transcribed into one polycistronic mRNA of 4.2 kb, and the transcription start was determined by the primer extension method to correspond with the first base of the ATG start codon of the sdhA gene. The S. acidocaldarius SdhA and SdhB subunits show characteristic sequence similarities to the succinate dehydrogenases and fumarate reductases of other organisms, while the SdhC and SdhD subunits, thought to form the membrane-anchoring domain, lack typical transmembrane ␣-helical regions present in all other succinate:quinone reductases (SQRs) and quinol:ifumarate reductases (QFRs) so far examined. Moreover, the SdhC subunit reveals remarkable 30% sequence similarity to the heterodisulfide reductase B subunit of Methanobacterium thermoautotrophicum and Methanococcus jannaschii, containing all 10 conserved cysteine residues. Electron paramagnetic resonance (EPR) spectroscopic studies of the purified enzyme as well as of membranes revealed the presence of typical S1 [2Fe2S] and S2 [4Fe4S] clusters, congruent with the deduced amino acid sequences. In contrast, EPR signals for a typical S3 [3Fe4S] cluster were not detected. However, EPR data together with sequence information implicate the existence of a second [4Fe4S] cluster in S. acidocaldarius rather than a typical [3Fe4S] cluster. These results and the fact that the S. acidocaldarius succinate dehydrogenase complex reveals only poor activity with caldariella quinone clearly suggest a unique structure for the SQR of S. acidocaldarius, possibly involving an electron transport pathway from the enzyme complex into the respiratory chain different from those for known SQRs and QFRs.Succinate:quinone reductase (SQR), a membrane-bound enzyme complex and component of the respiratory chain (complex II) of all aerobic organisms, catalyzes the oxidation of succinate to fumarate in the tricarboxylic acid cycle and transfers the electrons directly to quinones in the membrane. The reverse reaction, the reduction of fumarate, which is the last step in fumarate respiration of anaerobic or facultative anaerobic organisms, is catalyzed by quinol:fumarate reductase (QFR). The two enzyme complexes are very similar with regard to composition of subunits, prosthetic groups, and probably structure (2, 24). SQRs and QFRs are composed of three or four subunits of which the largest protein is a flavoprotein (M r of 65,000 to 79,000) carrying covalently bound 8-␣-N(3)-histidyl-flavin adenine dinucleotide (FAD). An iron-sulfur protein (M r of 25,000 to 37,000) together with this flavoprotein subunit represents the peripheral part of the enzyme complex. The iron-sulfur protein is suggested to contain three different types of iron-sulfur clusters, each equimolar with FAD. Magnetic circular dichroism and electron paramagnetic resonance...

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“…The first group of four conserved cysteinyl residues are the ligands for center 1, and this [2Fe-2S] cluster can form in the absence of the other clusters (4,16). The second domain encompasses the [4Fe-4S] and [3Fe-4S] clusters of FRD and SDH.…”

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Aerobic inactivation of fumarate reductase from Escherichia coli by mutation of the [3Fe-4S]-quinone binding domain

Cecchini¹,

Sices²,

Schröder³

et al. 1995

J Bacteriol

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Fumarate reductase from Escherichia coli functions both as an anaerobic fumarate reductase and as an aerobic succinate dehydrogenase. A site-directed mutation of E. coli fumarate reductase in which FrdB Pro-159 was replaced with a glutamine or histidine residue was constructed and overexpressed in a strain of E. coli lacking a functional copy of the fumarate reductase or succinate dehydrogenase complex. The consequences of these mutations on bacterial growth, assembly of the enzyme complex, and enzymatic activity were investigated. Both mutations were found to have no effect on anaerobic bacterial growth or on the ability of the enzyme to reduce fumarate compared with the wild-type enzyme. The FrdB Pro-159-to-histidine substitution was normal in its ability to oxidize succinate. In contrast, however, the FrdB Pro-159-to-Gln substitution was found to inhibit aerobic growth of E. coli under conditions requiring a functional succinate dehydrogenase, and furthermore, the aerobic activity of the enzyme was severely inhibited upon incubation in the presence of its substrate, succinate. This inactivation could be prevented by incubating the mutant enzyme complex in an anaerobic environment, separating the catalytic subunits of the fumarate reductase complex from their membrane anchors, or blocking the transfer of electrons from the enzyme to quinones. The results of these studies suggest that the succinate-induced inactivation occurs by the production of hydroxyl radicals generated by a Fenton-type reaction following introduction of this mutation into the [3Fe-4S] binding domain. Additional evidence shows that the substrate-induced inactivation requires quinones, which are the membrane-bound electron acceptors and donors for the succinate dehydrogenase and fumarate reductase activities. These data suggest that the [3Fe-4S] cluster is intimately associated with one of the quinone binding sites found in fumarate reductase and succinate dehydrogenase.During anaerobic respiration in Escherichia coli, the fumarate reductase (FRD) complex catalyzes the fumarate-dependent oxidation of menaquinol, thereby allowing production of a transmembrane proton gradient that can be used by the organism to support metabolism (19). The FRD complex is composed of four nonidentical subunits, FrdA, FrdB, FrdC, and FrdD, arranged into two domains: the FrdAB catalytic domain and the FrdCD membrane anchor domain (2, 22). FrdA (66 kDa) contains covalently bound flavin adenine dinucleotide and the active site of the enzyme (38), and the 27-kDa FrdB subunit contains the three distinct iron-sulfur centers present in the enzyme (23). The iron-sulfur clusters, center 1 ([2Fe-2S] ), are similar in structure and function in all membranebound succinate dehydrogenases (SDHs) and FRDs examined to date (2). The FrdC (15 kDa) and FrdD (13 kDa) subunits are essential for attachment of the catalytic subunits to the inner surface of the cytoplasmic membrane (22) and are also essential for electron transfer reactions involving menaquinol and ubiquinone (8).The p...

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Ligands to the 2Fe iron‐sulfur center in succinate dehydrogenase

Cited by 13 publications

References 37 publications

Studies on the assembly of complex II in yeast mitochondria using chimeric human/yeast genes for the iron-sulfur protein subunit

Studies on the assembly of complex II in yeast mitochondria using chimeric human/yeast genes for the iron-sulfur protein subunit

A succinate dehydrogenase with novel structure and properties from the hyperthermophilic archaeon Sulfolobus acidocaldarius: genetic and biophysical characterization

Aerobic inactivation of fumarate reductase from Escherichia coli by mutation of the [3Fe-4S]-quinone binding domain

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