Malate Dehydrogenases. I. A Survey of Molecular Size Measured by Gel Filtration

Murphey, William H.; Kitto, G. Barrie; Everse, Johannes; Kaplan, Nathan O.

doi:10.1021/bi00854a031

Cited by 113 publications

(44 citation statements)

References 26 publications

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“…The molecular weight (MW) of small MDHs is in the range from 60,000 to 74,000, and that of the large form is between 117,000 and 146,000. All MDHs studied so far are homogeneous in subunit composition, with a subunit MW in the range from 30,000 to 38,000 (1,16,17,23,30,31). The small enzymes are thus dimers, and the large enzymes are thus tetramers.…”

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confidence: 99%

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

1992

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Malate dehydrogenase (MDH; EC 1.1.1.37) from strain NCIB 8327 of the green sulfur bacterium Chlorobium vibrioforme was purified to homogeneity by triazine dye affinity chromatography followed by gel filtration. Purification of MDH gave an approximately 1,000-fold increase in specific activity and recoveries of typically 15 to 20%. The criteria of purity were single bands on sodium dodecyl sulfate (SDS) and nondenaturing polyacrylamide electrophoresis (PAGE) and the detection of a single N terminus in an Edman degradation analysis. MDH activity was detected in purified preparations by activity staining of gels in the direction of malate oxidation. PAGE and gel filtration (Sephadex G-100) analyses showed the native enzyme to be a dimer composed of identical subunits both at room temperature and at 4 degrees C. The molecular weight of the native enzyme as estimated by gel filtration was 77,000 and by gradient PAGE was 74,000. The subunit molecular weight as estimated by SDS-gradient PAGE was 37,500. N-terminal sequences of MDHs from C. vibrioforme, Chlorobium tepidum, and Heliobacterium gestii are presented. There are obvious key sequence similarities in MDHs from the phototrophic green bacteria. The sequences presented probably possess a stretch of amino acids involved in dinucleotide binding which is similar to that of Chloroflexus aurantiacus MDH and other classes of dehydrogenase enzymes but unique among MDHs.

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confidence: 99%

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

1992

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“…Based on molecular size, bacterial MDHs are divided into two groups: small and large (2,16,26). The molecular weight of small MDHs ranges from 60,000 to 65,000 (mammalian and other eucaryotic MDHs are in this category), and that of large MDHs is in the range of 117,000 to 146,000.…”

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confidence: 99%

Malate dehydrogenase in phototrophic purple bacteria: purification, molecular weight, and quaternary structure

Tayeh¹,

Madigan²

1987

J Bacteriol

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The citric acid cycle enzyme malate dehydrogenase was purified to homogeneity from the nonsulfur purple bacteria Rhodobacter capsulatus, Rhodospirillum rubrum, Rhodomicrobium vannielii, and Rhodocyclus purpureus. Malate dehydrogenase was purified from each species by either a single-or a two-step protocol; triazine dye affinity chromatography was the key step in purification of malate dehydrogenase in all cases. Purification of malate dehydrogenase resulted in a 130-to 240-fold increase in malate dehydrogenase specific activity, depending on the species, with recoveries ranging from 30 to 70%. Homogeneity of malate dehydrogenase preparations from the four organisms was determined by sodium dodecyl sulfate and nondenaturing polyacrylamide gel electrophoresis; a single protein band was observed in purified preparations by both techniques. The molecular weight of native malate dehydrogenases was determined by four independent methods and estimated to be in the range of 130,000 to 140,000 for the enzyme from R. capsulatus, R. rubrum, and R. vannielii and 57,000 for that from R. purpureus. It is concluded that malate dehydrogenase from R. capsulatus, R. rubrum, and R. vannielii is a tetramer composed of four identical subunits, while the enzyme from R. purpureus is a dimer composed of two identical subunits.

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“…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. …”

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confidence: 99%