An Apparent Oligomer of Malate Dehydrogenase from Bean Leaves

Habig, William H.; Racusen, David

doi:10.1104/pp.53.3.402

Cited by 15 publications

(7 citation statements)

References 51 publications

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“…The threedimensional structure of pig heart mitochondrial MDH has been determined by the X-ray-diffraction method [50]. The results indicated that the N-terminal sequence (residues 1-22) contains one fl-conformation (residues 1-5), one a-helix (residues [11][12][13][14][15][16][17][18][19][20][21][22] and one coenzymebinding domain (residues 6-10). The a-helix is found to reside in the interface between the subunits, and may be important for subunit-subunit interaction of the enzyme molecule.…”

Section: Km Valuesmentioning

confidence: 99%

“…In crude extracts it was shown that MDH might exist as oligomeric forms with the Mr much higher than the usual 60000-70000, as indicated by non-denaturing polyacrylamide/starch-gel electrophoresis and gel-filtration column chromatography [21]. These high-Mr MDH species, usually in addition to the normal-Mr form of the enzyme, have been reported in bacteria [1,2], fungi [6], animal mitochondria [22] and, most commonly, in higher plants [21,23,24]. These results indicated that the higherMr species of MDH in higher plants might result from aggregation of low-Mr species.…”

Section: Introductionmentioning

confidence: 99%

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Purification and molecular properties of malate dehydrogenase from the marine diatom Nitzschia alba

Yueh¹,

Chung²,

Lai³

1989

Biochemical Journal

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Malate dehydrogenase (EC 1.1.1.37) was purified to homogeneity from the marine diatom Nitzschia alba. The purification steps consisted of (NH4)2SO4 precipitation, ion-exchange chromatography, Blue Sepharose affinity chromatography and gel filtration. A typical procedure provided 685-fold purification with 58% yield. The Mr of the holoenzyme was estimated to be 322,000 by gel filtration and 316,000 by ultracentrifugation. The enzyme migrated as a single polypeptide spot on two-dimensional polyacrylamide-gel electrophoresis with an Mr of 38,500, suggesting that the holoenzyme consists of eight identical subunits. This is the first case where malate dehydrogenase has been shown to be a homo-octamer; malate dehydrogenases from other sources are predominantly homodimers, with two homotetramers reported so far. The amino acid composition of the enzyme was determined and the N-terminal sequence of the subunit polypeptide was found to be Arg-Lys-Val-Ala-Val-Met-Gly-Ala-Ala-Gly-Gly-Ile-Gly-Gln-Pro-Leu-Ser-Leu- Leu-Leu - Lys-Leu-Ser-Pro-Gln-Val-Thr-Glu-Leu-Ser-Lys-Tyr-. For the first 21 amino acid residues, near-identical sequences were reported for the enzymes isolated from pig heart, Escherichia coli, yeast and watermelon. Other physicochemical and catalytic properties, such as sedimentation coefficient, partial specific volume, Stokes radius, excitation and emission maxima, Michaelis constants, pH optima, pH stability range and activation energy, of this enzyme are also presented.

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Section: Km Valuesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Purification and molecular properties of malate dehydrogenase from the marine diatom Nitzschia alba

Yueh¹,

Chung²,

Lai³

1989

Biochemical Journal

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“…It now appears that previous studies on the aggregation behavior of MS under low-ionic-strength conditions overlooked its possible association with gMDH and that the decameric MS structure postulated by Henry et al (1992) is likely to correspond to an association between octameric MS and tetrameric gMDH. Our finding of MDH activity eluting as complexes of high Mr (670 kDa and 140 kDa) is not unusual; indeed, elution profiles of plant MDHs with apparent high-Mr values of 500kDa and 280kDa have been reported (O'Sullivan and Wedding 1972;Habig and Racusen 1974). Although the high-Mr MDH isoforms are very similar to the most commonly found low-Mr isoform with respect to optimal pH, K m for malate, and inhibition by various unreactive substrate analogs, they differ in their electrophoretic properties and ability to reduce 3-acetylpyridine-deamino-NAD + (Habig and Racusen 1974).…”

Section: Discussionmentioning

confidence: 64%

“…1A), and this indicates a possible aggregation between MS and an M D H isoform (coelution of a high-Mr MDH/MS complex was also observed on other columns with different separation ranges, including a Pharmacia Superose 6HR FPLC column and a Sepharose CL-4B column; data not shown). Plant MDH is usually resolved as a dimer (Mr 70000) with a subunit Mr of 35 000 (Habig and Racusen 1974). The higher Mr (140 000) estimated for MDH peak II ( Fig.…”

Section: Association~dissociation Of a Putative Ms/mdh Complexmentioning

confidence: 99%

Glyoxysomal malate dehydrogenase and malate synthase from soybean cotyledons (Glycine max L.): enzyme association, antibody production and cDNA cloning

Guex

Henry

Flach

et al. 1995

Planta

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In order to investigate a possible association between soybean malate synthase (MS; L-malate glyoxylate-lyase, CoA-acetylating, EC 4.1.3.2) and glyoxysomal malate dehydrogenase (gMDH; (S)-malate: NAD+ oxidoreductase, EC 1.1.1.37), two consecutive enzymes in the glyoxylate cycle, their elution profiles were analyzed on Superdex 200 HR fast protein liquid chromatography columns equilibrated in low- and high-ionic-strength buffers. Starting with soluble proteins extracted from the cotyledons of 5-d-old soybean seedlings and a 45% ammonium sulfate precipitation, MS and gMDH coeluted on Superdex 200 HR (low-ionic-strength buffer) as a complex with an approximate relative molecular mass (Mr) of 670,000. Dissociation was achieved in the presence of 50 mM KCl and 5 mM MgCl2, with the elution of MS as an octamer of M(r) 510,000 and of gMDH as a dimer of M(r) 73,000. Polyclonal antibodies raised to the native copurified enzymes recognized both denatured MS and gMDH on immunoblots, and their native forms after gel filtration. When these antibodies were used to screen a lambda ZAP II expression library containing cDNA from 3-d-old soybean cotyledons, they identified seven clones encoding gMDH, whereas ten clones encoding MS were identified using an antibody to SDS-PAGE-purified MS. Of these cDNA clones a 1.8 kb clone for MS and a 1.3-kb clone for gMDH were fully sequenced. While 88% identity was found between mature soybean gMDH and watermelon gMDH, the N-terminal transit peptides showed only 37% identity. Despite this low identity, the soybean gMDH transit peptide conserves the consensus R(X6)HL motif also found in plant and mammalian thiolases.

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“…The concentrated enzyme fractions were chromatographed on an UNO-Q column (7×35 mm, Bio-Rad), and the active fractions that were devoid of peroxidase activity were concentrated and used for MAD activity assays. The presence of MAD in the concentrated fractions was further verified by native polyacrylamide gel electrophoresis of the sample with subsequent substrate staining in a solution containing MTT, PMS, NAD + and malate [17].…”

Section: Detection Of Chorion Madmentioning

confidence: 99%

Chorion peroxidase-mediated NADH/O2 oxidoreduction cooperated by chorion malate dehydrogenase-catalyzed NADH production: a feasible pathway leading to H2O2 formation during chorion hardening in Aedes aegypti mosquitoes

Han¹,

Li²,

Li³

2000

Biochimica et Biophysica Acta (BBA) - General Subjects

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A specific chorion peroxidase is present in Aedes aegypti and this enzyme is responsible for catalyzing chorion protein cross-linking through dityrosine formation during chorion hardening. Peroxidase-mediated dityrosine cross-linking requires H 2 O 2 , and this study discusses the possible involvement of the chorion peroxidase in H 2 O 2 formation by mediating NADH/O 2 oxidoreduction during chorion hardening in A. aegypti eggs. Our data show that mosquito chorion peroxidase is able to catalyze pH-dependent NADH oxidation, which is enhanced in the presence of Mn 2+ . Molecular oxygen is the electron acceptor during peroxidase-catalyzed NADH oxidation, and reduction of O 2 leads to the production of H 2 O 2 , demonstrated by the formation of dityrosine in a NADH/peroxidase reaction mixture following addition of tyrosine. An oxidoreductase capable of catalyzing malate/NAD + oxidoreduction is also present in the egg chorion of A. aegypti. The cooperative roles of chorion malate/NAD + oxidoreductase and chorion peroxidase on generating H 2 O 2 with NAD + and malate as initial substrates were demonstrated by the production of dityrosine after addition of tyrosine to a reaction mixture containing NAD + and malate in the presence of both malate dehydrogenase fractions and purified chorion peroxidase. Data suggest that chorion peroxidase-mediated NADH/O 2 oxidoreduction may contribute to the formation of the H 2 O 2 required for chorion protein cross-linking mediated by the same peroxidase, and that the chorion associated malate dehydrogenase may be responsible for the supply of NADH for the H 2 O 2 production.

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An Apparent Oligomer of Malate Dehydrogenase from Bean Leaves

Abstract: ABSTRACT70,000 mol wt form of the enzyme. These higher mol wt species have been found in bacteria (38,39,56), fungi (5), higher plants (8,22,40,41,45,50), and animal mitochondria (10), usually in addition to the normal mol wt form of the enzyme.

Cited by 15 publications

References 51 publications

Purification and molecular properties of malate dehydrogenase from the marine diatom Nitzschia alba

Purification and molecular properties of malate dehydrogenase from the marine diatom Nitzschia alba

Glyoxysomal malate dehydrogenase and malate synthase from soybean cotyledons (Glycine max L.): enzyme association, antibody production and cDNA cloning

Chorion peroxidase-mediated NADH/O2 oxidoreduction cooperated by chorion malate dehydrogenase-catalyzed NADH production: a feasible pathway leading to H2O2 formation during chorion hardening in Aedes aegypti mosquitoes

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