Malate dehydrogenase from Chlorobium vibrioforme, Chlorobium tepidum, and Heliobacterium gestii: purification, characterization, and investigation of dinucleotide binding by dehydrogenases by use of empirical methods of protein sequence analysis

Charnock, Colin; Refseth, Unn Hilde; Sirevåg, Reidun

doi:10.1128/jb.174.4.1307-1313.1992

Cited by 25 publications

(13 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although RubisCO activity may be absent and the reductive carboxylic acid cycle is operative in C. tepidum, the latter metabolic pathway still requires participation of NAD(P)H in two steps of the cycle [6,7]: reduction of fumarate to malate by malate dehydrogenase [16] and the synthesis of isocitrate from succinate by reductive carboxylation catalyzed by isocitrate dehydrogenase [17].…”

Section: Introductionmentioning

confidence: 99%

Purification and characterization of ferredoxin–NAD(P)+ reductase from the green sulfur bacterium Chlorobium tepidum

Seo¹

2002

Biochimica et Biophysica Acta (BBA) - Protein Structure and Mol

View full text Add to dashboard Cite

Ferredoxin-NAD(P)+ reductase [EC 1.18 . It also reduces 2,6-dichlorophenol-indophenol (DPIP) and molecular oxygen with either NADPH or NADH as efficient electron donors. It showed NADPH diaphorase activity about two times higher than NADH diaphorase activity in DPIP reduction assays at NAD(P)H concentrations less than 0.1 mM. At 0.5 mM NAD(P)H, the two activities were about the same, and at 1 mM, the former activity was slightly lower than the latter.

show abstract

Section: Introductionmentioning

confidence: 99%

Purification and characterization of ferredoxin–NAD(P)+ reductase from the green sulfur bacterium Chlorobium tepidum

Seo¹

2002

Biochimica et Biophysica Acta (BBA) - Protein Structure and Mol

View full text Add to dashboard Cite

show abstract

“…This motif is, however, absent from the majority of L-MDHs investigated. It has been found only in L-MDHs from four eubacterial phototrophs, representing three phyla (2,19,25,30), and in two species of Bacillus (13, 38). Complete gene sequences are available for these L-MDHs (13,19,30,38), all of which show greater similarity in primary structure to L-lactate dehydrogenases (LLDHs) than to other L-MDHs.…”

mentioning

confidence: 99%

“…The resulting clear supernatant was the cell-free extract (CFE). L-MDH activity was measured as the reaction in the direction of oxaloacetate reduction by monitoring the oxidation of NADH at A 340 (2). An open glass column (inner diameter, 5 mm) containing approximately 1 ml of DyeMatrex gel Red-A (Amicon, Inc., Beverly, Mass.)…”

mentioning

confidence: 99%

Structural studies of malate dehydrogenases (MDHs): MDHs in Brevundimonas species are the first reported MDHs in Proteobacteria which resemble lactate dehydrogenases in primary structure

Charnock

1997

J Bacteriol

Self Cite

View full text Add to dashboard Cite

The N-terminal sequences of malate dehydrogenases from 10 bacterial strains, representing seven genera of Proteobacteria, were determined. Of these, the enzyme sequences of species classified in the genus Brevundimonas clearly resembled those malate dehydrogenases with greatest similarity to lactate dehydrogenases. Additional evidence from subunit molecular weights, peptide mapping, and enzyme mobilities suggested that malate dehydrogenases from species of the genus Brevundimonas were structurally distinct from others in the study.NAD-dependent L-malate dehydrogenase (L-malate-NADoxidoreductase, EC 1.1.1.37) (L-MDH) is an integral enzyme in several metabolic processes, including the tricarboxylic acid and glyoxylate cycles. In higher organisms, L-MDH isoenzymes are found in mitochondria, peroxisomes, and chloroplasts (NADP-dependent enzyme), in addition to the cytosol. The reaction catalyzed is the reversible oxidation of L-malate to oxaloacetate with NAD as the H acceptor. Although the oxidation of L-malate is in the usual physiological direction, in Chlorobium spp., which are obligate phototrophs, the reaction proceeds in the opposite direction as part of the reductive tricarboxylic cycle which fixes CO 2 (7).A stretch of glycine (G) residues has been implicated in the binding of NAD by most dehydrogenases with a requirement for this coenzyme (33). Three Gs lie separated as follows: GXGXXG, where X is any residue. This motif is, however, absent from the majority of L-MDHs investigated. It has been found only in L-MDHs from four eubacterial phototrophs, representing three phyla (2,19,25,30), and in two species of Bacillus (13, 38). Complete gene sequences are available for these L-MDHs (13,19,30,38), all of which show greater similarity in primary structure to L-lactate dehydrogenases (LLDHs) than to other L-MDHs. This similarity includes the possession of the putative NAD-binding configuration GXGXXG by both dehydrogenases. Using data derived from empirical methods of protein sequence analysis and existing X-ray crystallography data, we have previously proposed an alternative G motif from that employed by most L-MDHs, i.e., GAXGXXG/A, where A is alanine (2). L-MDH shares many structural and catalytic features with L-LDH which are suggestive of a common ancestry (1,10,17). The existence of LMDHs which most resemble L-LDHs in primary structure, and their distribution in the bacterial phyla, may offer clues about the ancestral links between these two classes of dehydrogenase enzymes. The NAD-binding motif of L-MDHs is located close to the N terminus. Consequently, sequencing of this region of the enzyme provides data of particular value for comparing L-MDHs with each other and with L-LDHs. Relatively few sequences have been determined for species of Proteobacteria (16,26,32), and to date, all those investigated possess the GAXGXXG/A motif.Isolation of L-MDH from select Proteobacteria. Two liters of Luria broth were seeded with two to three bacterial colonies (strains used in the study are listed in Table 1), and ...

show abstract

“…In LDH, this is GXGXXG, whereas for most known MDHs, it is GAXGXX(G/A). However, MDHs from all photosynthetic bacteria studied so far have the glycine motif typical of LDH (4,29).…”

mentioning

confidence: 99%

“…as the standard. MDH (L-malate:NAD ϩ oxidoreductase; EC 1.1.1.37) activity was assayed as described by Charnock et al (4). To acertain that the purified MDHs were equally free of NAD(H), assays were performed without the presence of cofactor.…”

mentioning

confidence: 99%

Malate dehydrogenase from the mesophile Chlorobium vibrioforme and from the mild thermophile Chlorobium tepidum: molecular cloning, construction of a hybrid, and expression in Escherichia coli

1996

Self Cite

View full text Add to dashboard Cite

The genes (mdh) encoding malate dehydrogenase (MDH) from the mesophile Chlorobium vibrioforme and the moderate thermophile C. tepidum were cloned and sequenced, and the complete amino acid sequences were deduced. When the region upstream of mdh was analyzed, a sequence with high homology to an operon encoding ribosomal proteins from Escherichia coli was found. Each mdh gene consists of a 930-bp open reading frame and encodes 310 amino acid residues, corresponding to a subunit weight of 33,200 Da for the dimeric enzyme. The amino acid sequence identity of the two MDHs is 86%. Homology searches using the primary structures of the two MDHs revealed significant sequence similarity to lactate dehydrogenases. A hybrid mdh was constructed from the 3 part of mdh from C. tepidum and the 5 part of mdh from C. vibrioforme. The thermostabilities of the hybrid enzyme and of MDH from C. vibrioforme and C. tepidum were compared.Malate dehydrogenase (MDH) catalyzes the reversible conversion of malate to oxaloacetate, using NAD as cofactor. In most organisms, MDH is an essential enzyme in carbon metabolism, and in green sulfur bacteria, it is part of the reductive tricarboxylic acid cycle (9).Complete amino acid sequence data for MDH are available from many species within the Bacteria, Archaea, and Eucarya. Based on sequence similarity, Goward and Nicholls (15) have suggested that MDH has diverged into two distinct phylogenetic groups, one group that includes cytoplasmic MDH, chloroplast MDH, and MDH from Thermus flavus, and a second group that includes MDHs with similarity to lactate dehydrogenase (LDH). In the latter group, the mitochondrial MDH and eubacterial MDH are in one cluster and the more LDHlike (archaeal) Haloarcula marismortui MDH is in another.Although the primary structures of the various MDHs and LDHs are different, their tertiary structures are similar. The dehydrogenases have two domains; the structurally conserved N-terminal dinucleotide-binding domain, and the C-terminal catalytic domain, which differs among the different dehydrogenases. The dinucleotide-binding domain of the LDHs and MDHs that use NAD as a cofactor contains a characteristic glycine motif. In LDH, this is GXGXXG, whereas for most known MDHs, it is GAXGXX(G/A). However, MDHs from all photosynthetic bacteria studied so far have the glycine motif typical of LDH (4, 29).The amino acid residues important for substrate and coenzyme binding, catalysis, and subunit assembly have been identified in several MDHs and LDHs (3,27,36,42). With regard to thermostability, LDH from Bacillus species has been extensively studied (21,41,45,46), and studies have also been performed with MDH from Escherichia coli (14) and from the thermophilic Thermus aquaticus (6).In the present work, MDH and its corresponding gene from the two green sulfur bacteria Chlorobium tepidum and Chlorobium vibrioforme have been isolated, characterized, and compared. The green sulfur bacteria comprise a distinct phylogenetic group of photosynthetic species, which are related to other gro...

show abstract

Malate dehydrogenase from Chlorobium vibrioforme, Chlorobium tepidum, and Heliobacterium gestii: purification, characterization, and investigation of dinucleotide binding by dehydrogenases by use of empirical methods of protein sequence analysis

Cited by 25 publications

References 44 publications

Purification and characterization of ferredoxin–NAD(P)+ reductase from the green sulfur bacterium Chlorobium tepidum

Purification and characterization of ferredoxin–NAD(P)+ reductase from the green sulfur bacterium Chlorobium tepidum

Structural studies of malate dehydrogenases (MDHs): MDHs in Brevundimonas species are the first reported MDHs in Proteobacteria which resemble lactate dehydrogenases in primary structure

Malate dehydrogenase from the mesophile Chlorobium vibrioforme and from the mild thermophile Chlorobium tepidum: molecular cloning, construction of a hybrid, and expression in Escherichia coli

Contact Info

Product

Resources

About