Isolation and characterization of bovine lactate dehydrogenase x

Kolb, E; Fleisher, Gerard A.; Larner, Joseph

doi:10.1021/bi00824a018

Cited by 32 publications

(2 citation statements)

References 28 publications

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“…LDH-C 4 catalyzes the interconversion of pyruvate to lactate with the concomitant oxidation of NADH to NAD + , which is essential for the continued production of Adenosine Triphosphate (ATP) by glycolysis [ 34 ]. LDH-C 4 has unique structural and functional properties [ 35 , 36 , 37 ].Comparative studies of catalytic properties have revealed differences between LDH-C 4 and other lactate dehydrogenase isozymes [ 38 , 39 ] LDH-C 4 from different species shows activity against α-keto and α-hydroxy acids of longer carbon chains than those of pyruvate and lactate [ 35 , 36 , 37 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 ]. Substrate specificity differences between LDH isozymes and LDH-C 4 isozymes from different species suggest that N-substituted oxamates are strong inhibitors of LDH-C 4 activity [ 48 ].…”

Section: Introductionmentioning

confidence: 99%

Testis-Specific Lactate Dehydrogenase (LDH-C4) in Skeletal Muscle Enhances Apika’s Sprint-Running Capacity in Hypoxic Environment

Wang

Lian

Wei

et al. 2015

IJERPH

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LDH-C4 is a lactate dehydrogenase that catalyzes the conversion of pyruvate to lactate. In mammals, ldh-c was originally thought to be expressed only in testis and spermatozoa. Plateau pika (Ochotona curzoniae), which belongs to the genus Ochotona of the Ochotonidea family, is a hypoxia tolerant mammal living 3000–5000 m above sea level on the Qinghai-Tibet Plateau, an environment which is strongly hypoxic. Ldh-c is expressed not only in testis and sperm but also in somatic tissues of plateau pika. In this study, the effects of N-propyl oxamate and N-isopropyl oxamate on LDH isozyme kinetics were compared to screens for a selective inhibitor of LDH-C4. To reveal the role and physiological mechanism of LDH-C4 in skeletal muscle of plateau pika, we investigated the effect of N-isopropyl oxamate on the pika exercise tolerance as well as the physiological mechanism. Our results show that Ki of N-propyl oxamate and N-isopropyl oxamate for LDH-A4, LDH-B4, and LDH-C4 were 0.094 mmol/L and 0.462 mmol/L, 0.119 mmol/L and 0.248 mmol/L, and 0.015 mmol/L and 0.013 mmol/L, respectively. N-isopropyl oxamate is a powerful selective inhibitor of plateau pika LDH-C4. In our exercise tolerance experiment, groups treated with inhibitors had significantly lower swimming times than the uninhibited control group. The inhibition rates of LDH, LD, and ATP were 37.12%, 66.27%, and 32.42%, respectively. Our results suggested that ldh-c is expressed in the skeletal muscle of plateau pika, and at least 32.42% of ATP in the skeletal muscle is catalyzed by LDH-C4 by anaerobic glycolysis. This suggests that pika has reduced dependence on oxygen and enhanced adaptation to hypoxic environment due to increased anaerobic glycolysis by LDH-C4 in skeletal muscle. LDH-C4 in plateau pika plays the crucial role in anaerobic glycolysis and generates ATP rapidly since this is the role of LDH-A4 in most species on plain land, which provide evidence that the native humans and animals in Qinghai-Tibet plateau can adapt to the hypoxia environment.

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Section: Introductionmentioning

confidence: 99%

Testis-Specific Lactate Dehydrogenase (LDH-C4) in Skeletal Muscle Enhances Apika’s Sprint-Running Capacity in Hypoxic Environment

Wang

Lian

Wei

et al. 2015

IJERPH

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“…LDH-C 4 has high thermostability [ 17 ], and its homolog from different species showed activity against longer carbon chain with α-hydroxy and α-keto acids than those of pyruvate and lactate [ 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 ], indicating that LDH-C 4 has unique structural and functional properties [ 31 , 40 , 41 , 42 ]. In the present study, we compared the enzymatic kinetic characteristics among LDH-A 4 , LDH-B 4 , and LDH-C 4 of plateau pika with the aim of exploring the pika’s adaptation mechanism to the hypoxic environment of the Qinghai-Tibet Plateau.…”

Section: Introductionmentioning

confidence: 99%

Enzymatic Kinetic Properties of the Lactate Dehydrogenase Isoenzyme C4 of the Plateau Pika (Ochotona curzoniae)

Wang¹,

Lian²,

Wei³

et al. 2016

IJMS

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Testis-specific lactate dehydrogenase (LDH-C4) is one of the lactate dehydrogenase (LDH) isozymes that catalyze the terminal reaction of pyruvate to lactate in the glycolytic pathway. LDH-C4 in mammals was previously thought to be expressed only in spermatozoa and testis and not in other tissues. Plateau pika (Ochotona curzoniae) belongs to the genus Ochotona of the Ochotonidea family. It is a hypoxia-tolerant species living in remote mountain areas at altitudes of 3000–5000 m above sea level on the Qinghai-Tibet Plateau. Surprisingly, Ldh-c is expressed not only in its testis and sperm, but also in somatic tissues of plateau pika. To shed light on the function of LDH-C4 in somatic cells, Ldh-a, Ldh-b, and Ldh-c of plateau pika were subcloned into bacterial expression vectors. The pure enzymes of Lactate Dehydrogenase A4 (LDH-A4), Lactate Dehydrogenase B4 (LDH-B4), and LDH-C4 were prepared by a series of expression and purification processes, and the three enzymes were identified by the method of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and native polyacrylamide gel electrophoresis (PAGE). The enzymatic kinetics properties of these enzymes were studied by Lineweaver-Burk double-reciprocal plots. The results showed the Michaelis constant (Km) of LDH-C4 for pyruvate and lactate was 0.052 and 4.934 mmol/L, respectively, with an approximate 90 times higher affinity of LDH-C4 for pyruvate than for lactate. At relatively high concentrations of lactate, the inhibition constant (Ki) of the LDH isoenzymes varied: LDH-A4 (Ki = 26.900 mmol/L), LDH-B4 (Ki = 23.800 mmol/L), and LDH-C4 (Ki = 65.500 mmol/L). These data suggest that inhibition of lactate by LDH-A4 and LDH-B4 were stronger than LDH-C4. In light of the enzymatic kinetics properties, we suggest that the plateau pika can reduce reliance on oxygen supply and enhance its adaptation to the hypoxic environments due to increased anaerobic glycolysis by LDH-C4.

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Marsupial and monotreme lactate dehydrogenase isozymes: Phylogeny, ontogeny, and homology eutherian mammals

1973

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Lactate dehydrogenase (LDH) isozymes have been examined from tissues of mammalian species representative of the five families of marsupials and of the monotremes. Immunochemical, molecular hybridization, and tissue distribution studies showed that three genes (A, B and C) are responsible for LDH synthesis giving rise to five major isozymes in most tissues (LDHs &, A&, A2B2, A1B3, and B4) and an additional isozyme (LDH-C4) which is present only in mature testes. The LDH phenotypes of 93 erythrocyte samples from 31 marsupial and 2 monotreme species were analysed. Although the isozymic composition varied markedly within the marsupial families Macropodidae and Phalangeridae, the Dasyuridae showed remarkable uniformity, containing only LDH-b. The two monotreme species analysed also exhibited this latter phenotype in their erythrocytes. Ontogenic studies of the LDH isozymes of pouch young from Macropus parryi (pretty-face wallaby) showed changes similar to those reported for the developing mammalian fetus in utero. The immature pouch young exhibited similar tissue patterns of LDH isozymes which diverged during growth in the pouch thus reflecting differential genetic activity. A comparison of the relative maturity of the eutherian fetus with marsupial pouch young may be made. From these observations, it is concluded that a high degree of correlation exists among the properties of eutherian, marsupial, and monotreme LDH isozymes.

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Isolation and characterization of bovine lactate dehydrogenase x

Cited by 32 publications

References 28 publications

Testis-Specific Lactate Dehydrogenase (LDH-C4) in Skeletal Muscle Enhances Apika’s Sprint-Running Capacity in Hypoxic Environment

Testis-Specific Lactate Dehydrogenase (LDH-C4) in Skeletal Muscle Enhances Apika’s Sprint-Running Capacity in Hypoxic Environment

Enzymatic Kinetic Properties of the Lactate Dehydrogenase Isoenzyme C4 of the Plateau Pika (Ochotona curzoniae)

Marsupial and monotreme lactate dehydrogenase isozymes: Phylogeny, ontogeny, and homology eutherian mammals

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