Processing of <i>Bacillus subtilis</i> succinate dehydrogenase and cytochrome &amp;‐558 polypeptides

Hederstedt, Lars; Bergman, Tomas; Jörnvall, Hans

doi:10.1016/0014-5793(87)81527-2

Cited by 30 publications

(18 citation statements)

References 26 publications

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“…To what extent the reaction leading to the covalent flavinylation of other enzymes [22] shows similarities to the reaction described here for 6-HDNO remains to be seen.…”

Section: Discussionmentioning

confidence: 88%

Phosphoenolpyruvate‐dependent flavinylation of 6‐hydroxy‐D‐nicotine oxidase

Nagursky

Bichler

Brandsch

1988

European Journal of Biochemistry

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The reaction leading to the flavinylation of apo-6-hydroxy-~-nicotine oxidase was investigated in cell-free extracts of Eschericia coli carrying the 6-hydroxy-~-nicotine oxidase (6-HDNO) gene on the expression plasmid pDB222. It was demonstrated that the reaction required phosphoenolpyruvate (P-pyruvate) in addition to FAD. When [32P]P-pyruvate or [14C]P-pyruvate were used in the reaction with apo-6-HDN0, no phosphorylated or pyruvylated apo-protein could be detected, however. In order to drive the reaction to completion, FAD and Ppyruvate had to be present simultaneously in the reaction mixture. When apo-6-HDN0, highly purified by affinity chromatography, was used in the reaction with P-pyruvate and FAD, no additional protein fraction was required. A possible reaction scheme for the formation of holoenzyme from 6-HDNO is discussed.Enzymes which are covalently modified by the attachment of their cofactor to the side chain of a specific amino acid residue represent a special category of post-translational modifications. The functional significance of these modifications and how they are accomplished is poorly understood. The covalent attachment of biotin to a specific lysine residue of carboxylases [l] and of heme to two specific cysteine residues of cytochrome c [2] are known to take place enzymatically. No corresponding holoenzyme synthetase, however, has been described so far for enzymes carrying covalently bound lipoic acid or covalently bound flavin. Both flavin cofactors, FAD as well as FMN, have been described as occurring in covalent linkage [3]. Biotin and lipoic-aciddependent enzymes always contain the cofactor bound to the polypeptide chain by a specific lysine residue [4]. Among the cytochromes, only cytochromes c andfcarry heme [5], always covalently bound by two essential cysteine residues. The situation with FAD, the primary flavin cofactor, is different. This cofactor is tightly but noncovalently bound in the majority of enzymes. In different covalently flavinylated enzymes, FAD is bound to the side chain of different amino acid residues. The 8a-methyl group of the FAD isoaloxazine ring is linked to the N1 or N3 of a histidine, to the sulfur of a cysteine or the oxygen of a tyrosine residue; 6-S-cysteinyl-FMN was also identified in some enzymes (reviewed in [3]). In addition to the differences in type of FAD linkage to the protein moiety, no amino acid sequence similarities surrounding the amino acid residue involved in FAD binding have been found thus far between covalently flavinylated enzymes. This situation is in contrast to information obtained from other examples of covalent modification of enzymes by cofactors where similar binding sequences are present [2, 3, 61. The question of whether the same mechanism accounts for €he covalent attachCorrespondence to R. Brandsch, Biochemisches Institut, Universit i t Freiburg, Hermann-Herder-StraBe 7, D-7800 Freiburg, Federal Republic of Germany Abbreviations. 6-HDNO, 6-hydroxy-~-nicotine oxidase; P-pyruvate, phosphoenolpyruvate.Enzyme. 6-Hydroxy-~-ni...

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“…To what extent the reaction leading to the covalent flavinylation of other enzymes [22] shows similarities to the reaction described here for 6-HDNO remains to be seen.…”

Section: Discussionmentioning

confidence: 88%

Phosphoenolpyruvate‐dependent flavinylation of 6‐hydroxy‐D‐nicotine oxidase

Nagursky

Bichler

Brandsch

1988

European Journal of Biochemistry

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“…The N-terminal end of the bacterial polypeptides function as signal peptides [12] but are essentially not processed, i.e. four residues are removed from E. coli SdhC [21] and the initiating-methionine in B. subtilis SdhC [22] is removed in about 50% of the cases. (iv) Proline and glycine residues are frequent in the hydrophilic segments, supporting the view that these parts are structurally flexible to function as connecting loops.…”

Section: The Cytoehrome B Componentmentioning

confidence: 99%

“…The deduced polypeptide sequences are very similar to E. eoli FrdCD and Bos taurus SdhC, respectively, and were therefore not included in the alignment. The N-terminus of the BsS [22], BoS [18], and ScS SdhC [20] and SdhD [19] polypeptides as shown in Fig. 3 is that determined by analysis of polypeptide isolated from the respective enzyme.…”

Section: A 3-dimensional Structural Modelmentioning

confidence: 99%

A structural moDAl for the membrane‐integral domain of succinate:quinone oxidoreductases

Hägerhäll¹,

Hederstedt²

1996

FEBS Letters

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123

102

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Many succinate:quinone oxidoreductases in bacteria and mitochondria, i.e. succinate:quinone reductases and fumarate reductases, contain in the membrane anchor a cytochrome b whose structure and function is poorly understood. Based on biochemical data and polypeptide sequence information, we show that the anchors in different organisms are related despite an apparent diversity in polypeptide and heine composition. A general structural model for the membrane-integral domain of the anchors is proposed. It is an antiparallel four-helix bundle with a novel arrangement of hexa-coordinated protoheme IX. The structure can be applied to a larger group of membraneintegral cytochromes of b-type and has evolutionary and functional implications.

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“…The data were taken to indicate that the correct formation of a binding site for FAD by residues throughout the peptide chain is necessary for flavinylation to proceed. A requirement for cell-specific factors for FAD attachment was demonstrated by the expression of the B. subtilis sdh operon in E. coli (Hederstedt et al, 1987). Heterologous expression in E. coli leads to the flavoprotein subunit lacking covalently bound FAD, and being incorrectly processed at the N-terminus.…”

mentioning

confidence: 99%

Covalent attachment of flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN) to enzymes: The current state of affairs

1998

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The first identified covalent flavoprotein, a component of mammalian succinate dehydrogenase, was reported 42 years ago. Since that time, more than 20 covalent flavoenzymes have been described, each possessing one of five modes of FAD or FMN linkage to protein. Despite the early identification of covalent flavoproteins, the mechanisms of covalent bond formation and the roles of the covalent links are only recently being appreciated. The main focus of this review is, therefore, one of mechanism and function, in addition to surveying the types of linkage observed and the methods employed for their identification. Case studies are presented for a variety of covalent flavoenzymes, from which general findings are beginning to emerge.Keywords: covalent flavoproteins; flavin adenine dinucleotide; flavin mononucleotide; flavinylation; redox cofactors Cofactors are recruited by protein molecules to extend the chemistry available within the active sites of enzymes. The chemistries displayed are variable and the range of protein-specific cofactors available bears testimony to the types of biological reaction achieved by many enzyme molecules. Cofactors may be present in the freely dissociable form, but others, such as lipoic acid and biotin, are permanently linked to the host protein. Other cofactors (e.g., heme and pyridoxyl phosphate) fall into both categories in that they can be attached covalently to the protein or operate in the freely dissociable form. The covalent addition of specific cofactors to unique residues within a polypeptide chain is a fundamental problem in studies of biological molecular recognition. In many cases, this specificity is purchased at the expense of recruiting modification enzymes that direct the covalent addition of a cofactor to the host protein. For example, the additions of biotin and lipoic acid to the €-amino groups of specific Lys residues in carboxyltransferases and the 2-oxoacid dehydrogenases are directed by a biotin holoenzyme synthase (Sweetman et al., 1985;Takai et al., 1987) and Reprint requests to: N.S. Scmtton, Department of Biochemistry, University of Leicester, Adrian Building, University Road, Leicester LE1 7RH UK; e-mail: nss4@le.ac.uk; or W.S. McIntire, Molecular Biology Division, Department of Veterans Affairs Medical Center, San Francisco, California 94121; e-mail: wsm@itsa.ucsf.edu.lipoate-protein ligases (Brookfield et al., 1991; Moms et al., 1994), respectively. In a similar fashion, a heme cytochrome c lyase (Steiner et al., 1996) is responsible for the covalent addition of heme to two Cys residues via thioether linkages. In some enzymes, an existing side chain is modified to yield what is in effect a covalently attached cofactor, although no pre-existing cofactor molecule is incorporated into the enzyme. Several examples of this type of modification are provided in the next paragraph.For histidine decarboxylase of Lactobacillus 30A (Recsei & Snell, 1984), a pyruvoyl group is generated from an existing residue, Ser 82. The enzyme is synthesized as a proenzym...

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Processing of Bacillus subtilis succinate dehydrogenase and cytochrome &‐558 polypeptides

Cited by 30 publications

References 26 publications

Phosphoenolpyruvate‐dependent flavinylation of 6‐hydroxy‐D‐nicotine oxidase

Phosphoenolpyruvate‐dependent flavinylation of 6‐hydroxy‐D‐nicotine oxidase

A structural moDAl for the membrane‐integral domain of succinate:quinone oxidoreductases

Covalent attachment of flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN) to enzymes: The current state of affairs

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