The preparation of the cytoplasmic and mitochondrial forms of malate dehydrogenase and aspartate aminotransferase from pig heart by a single procedure

Glatthaar, Beat E.; Barbarash, Gary R.; Noyes, Barbara E.; Banaszak, Leonard J.; Bradshaw, Ralph A.

doi:10.1016/0003-2697(74)90099-2

Cited by 64 publications

(19 citation statements)

References 29 publications

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“…With the exception of the composition reported by Anderton (20), the values obtained with the homogeneous protein (column 4) are in close agreement with the preparations that also contain the more acidic subforms (columns 1 and 3). This finding is consistent with previous observations (14,22 (19) and in quantitative agreement with a dimeric subunit structure (26).…”

Section: Characterization Of the Principal Component Of Pig Heartsupporting

confidence: 93%

“…The isoelectric point, determined from the measured pH of the eluted fractions, is 9.3. This value is entirely consistent with the observed behavior of the enzyme on CM-cellulose and starch gel electrophoresis (14,27 (29). In subsequent conversion of the tritiated NADH to NAD+ by L-3-hydroxyacyl CoA dehydrogenase or mitochondrial malate dehydrogenase, the tritium should remain in the NAD+ if the enzyme is A-specific, but should be transferred to the substrate if the enzyme is Bspecific.…”

Section: Characterization Of the Principal Component Of Pig Heartsupporting

confidence: 90%

“…EXPERIMENTAL PROCEDURE Homogeneous L-3-hydroxyacyl CoA dehydrogenase and mitochondrial malate dehydrogenase from pig heart muscle were prepared as described previously (13,14). In each case, the principal component was isolated free of the more acidic subforms that have been present in the preparations of other workers.…”

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confidence: 99%

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Structural and Functional Similarities Between Mitochondrial Malate Dehydrogenase and L-3-Hydroxyacyl CoA Dehydrogenase

Noyes

Glatthaar

Garavelli

et al. 1974

Proc. Natl. Acad. Sci. U.S.A.

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Pig heart mitochondrial malate dehydrogenase (EC 1.1.1.37), which has been obtained free of electrophoretic subforms, has been shown to have a molecular weight of 67,000 and to be composed of two polypeptide chains. Comparison of these and other properties, such as amino-acid composition, isoelectric point, and keto-substrate inhibition, with those of L-3-hydroxyaeyl CoA dehydrogenase (EC 1.1.1.35), another NAD+-dependent dehydrogenase of mitochondrial origin, suggests structural similarities of the type associated with proteins possessing common evolutionary origins. This conclusion is supported by immunological crossreactivity. In view of these observations, the dissimilarity in the stereospecificity of hydrogen transfer from cofactor to substrate catalyzed by the two enzymes is attributed to 1800 rotation -in the binding orientation of the nicotinamide moiety of the NAD+, rather than to gross differences in the geometry of the active site of the two enzymes.Comparisons of the structure-function relationships of numerous proteins have provided a clearly emerging picture of the evolution of biological function within a class of proteins through selective genetic alterations leading to local changes in side chain character and environment without appreciably affecting the conformation of the polypeptide backbone. The family of serine proteases, where similarities in primary and three-dimensional structure are manifested in a variety of proteolytic enzymes with differing specificities from diverse phylogenetic origins (1)(2)(3)(4)(5), is the best documented example of this phenomenon. Altho~h less well documented at the threedimensional level, Iyaouyme and a-lactalbumin (6, 7) and nerve growth factor and insulin (8)

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Section: Characterization Of the Principal Component Of Pig Heartsupporting

confidence: 93%

Section: Characterization Of the Principal Component Of Pig Heartsupporting

confidence: 90%

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Structural and Functional Similarities Between Mitochondrial Malate Dehydrogenase and L-3-Hydroxyacyl CoA Dehydrogenase

Noyes

Glatthaar

Garavelli

et al. 1974

Proc. Natl. Acad. Sci. U.S.A.

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“…Sci. USA 84 (1987) 8263 previously described (8,9). In the low-resolution study, three heavy-atom derivatives were used for phase determination, and two of these, methylmercury(II) chloride and K2PtCl6 were usable for the high-resolution work.…”

Section: Methodsmentioning

confidence: 99%

Structure of L-3-hydroxyacyl-coenzyme A dehydrogenase: preliminary chain tracing at 2.8-A resolution.

Birktoft

Holden²,

Hamlin³

et al. 1987

Proc. Natl. Acad. Sci. U.S.A.

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The conformation of L-3-hydroxyacyl-CoA dehydrogenase (EC 1.1.1.35) has been derived from electrondensity maps calculated at 2.8-A resolution with phases obtained from two heavy-atom derivatives and the bound coenzyme, NAD. Like other dehydrogenases, 3-hydroxyacyl-CoA dehydrogenase is a double-domain structure, but the bilobal nature of this enzyme is more pronounced than has been previously observed. The amino-terminal domain, which comprises approximately the first 200 residues, is responsible for binding the NAD cofactor and displays considerable structural homology with the dinucleotide binding domains observed in other NAD-, NADP-, and FAD-dependent enzymes. The carboxyl-terminal domain, comprising the remaining 107 residues, appears to be all a-helical and bears little homology to other known dehydrogenases. The subunit-subunit interface in the 3-hydroxyacyl-CoA dehydrogenase dimer is formed almost exclusively by residues in the smaller helical domain. A difference map between the apo and holo forms of the crystalline enzyme has been interpreted in terms of the NAD molecule being bound in a typically extended conformation. The location of the coenzyme binding site, along with the structural homology to other dehydrogenases, makes it possible to speculate about the location of the binding site for the fatty acyl-CoA substrate.In most organisms, the metabolic breakdown of long-chain fatty acids proceeds through the,B-oxidation pathway. In this process, a thioester is formed between CoA and the fatty acid, and the alkyl chain is then degraded by the sequential removal of two-carbon units. The mitochondrial a-oxidation cycle involves four enzymes; one of these is L-3-hydroxyacyl-CoA dehydrogenase [(S)-3-hydroxyacyl-CoA:NAD+ oxidoreductase, EC 1.1.1.35]. This enzyme utilizes NAD as a cofactor and L-3-hydroxyacyl-CoA as substrate in the following reaction, which in vivo proceeds exclusively to the right:

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“…Lactate dehydrogenase (LDH-I-L,) and cytoplasmatic malate dehydrogenase (s-MDH) were prepared from pig heart by slight modifications of published procedures [12,13] and their activity assayed as described therein.…”

Section: Enzymesmentioning

confidence: 99%

Phenol‐containing adsorbents for the affinity chromatography purification of some NAD(H)‐dependent dehydrogenases

Brenna¹,

Roux²,

Pietta³

et al. 1977

FEBS Letters

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The preparation of the cytoplasmic and mitochondrial forms of malate dehydrogenase and aspartate aminotransferase from pig heart by a single procedure

Cited by 64 publications

References 29 publications

Structural and Functional Similarities Between Mitochondrial Malate Dehydrogenase and L-3-Hydroxyacyl CoA Dehydrogenase

Structural and Functional Similarities Between Mitochondrial Malate Dehydrogenase and L-3-Hydroxyacyl CoA Dehydrogenase

Structure of L-3-hydroxyacyl-coenzyme A dehydrogenase: preliminary chain tracing at 2.8-A resolution.

Phenol‐containing adsorbents for the affinity chromatography purification of some NAD(H)‐dependent dehydrogenases

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