Browse > Article
http://dx.doi.org/10.5352/JLS.2009.19.2.256

Purification and Characterization of Lactate Dehydrogenase A4 Isozyme in Mandrin Fish (Siniperca scherzeri)  

Cho, Sung-Kyu (Industrial Science Research Institute, Cheongju University)
Ku, Bo-Ra (Department of Life Science, Cheongju University)
An, Hyo-Jung (Department of Life Science, Cheongju University)
Park, Eun-Mi (Department of Life Science, Cheongju University)
Park, Seon-Young (Department of Life Science, Cheongju University)
Kim, Jae-Bum (Department of Anatomy, School of Medicine, Sungkyunkwan University)
Yum, Jung-Joo (Department of Life Science, Cheongju University)
Publication Information
Journal of Life Science / v.19, no.2, 2009 , pp. 256-263 More about this Journal
Abstract
The lactate dehydrogenase (EC 1.1.1.27, LDH) $A_4$ isozyme in skeletal muscle of mandrin fish (Siniperca scherzeri) was successfully purified by affinity chromatography and ultrafiltration. The molecular weight of the purified LDH $A_4$ isozyme was 140.4 kDa and its isoelectric point (pI) was 7.0. Optimal pH for enzymatic reaction was 7.5. ${K_m}^{PYR}$ and $V_{max}$ value of the purified LDH $A_4$ isozyme were $4.86{\times}10^{-5}$ M and 13.31 mM/min using pyruvate as a substrate, respectively. These kinetic properties of the purified LDH $A_4$ isozyme supported the fact that the mandrin fish was a warm-adapted species. The antibody against the purified LDH $A_4$ isozyme may be used in the metabolic physiological studies of ectothermic vertebrates and in the diagnosis of several human diseases.
Keywords
Mandrin fish (Siniperca scherzeri); lactate dehydrogenase (LDH); $A_4$ isozyme; purification; antibody;
Citations & Related Records
Times Cited By KSCI : 3  (Citation Analysis)
연도 인용수 순위
1 Sevinc, A., R. Sari, and E. Fadillioglu. 2005. The utility of lactate dehydrogenase isoenzyme pattern in the diagnostic evaluation of malignant and nonmalignant ascites. J. Natl. Med. Assoc. 97, 79-84
2 Sidell, B. D. and K. F. Beland. 1980. Lactate dehydrogenases of Atlantic hagfish: physiological and evolutionary implications of a primitive heart isozyme. Science 207, 769-770   DOI
3 Whitt, G. S. 1970. Developmental genetics of the lactate dehydrogenase isozymes of fish. J. Exp. Zool. 175, 1-35   DOI   ScienceOn
4 Yancey, P. H. and J. F. Siebenaller. 1987. Coenzyme binding ability of homologs of M4-lactate dehydrogenase in temperature adaptation. Biochim. Biophys. Acta 924, 483-491   DOI   ScienceOn
5 Yum, J. J. 2008. Characterization of lactate dehydrogenase in Acanthogobius hasta. J. Life Sci. 18, 264-272   과학기술학회마을   DOI
6 Yum, J. J. and M. O. Kim. 1989. Biochemical properties of lactate dehydrogenase isozymes in Pseudogobio esocinus. J. Ind. Sci., Cheongju Univ. Korea 7, 151-162
7 Markert, C. L., J. B. Shaklee, and G. S. Whitt. 1975. Evolution of a gene: multiple genes for LDH isozymes provide a model of the evolution of gene structure, function and regulation. Science 189, 102-114   DOI   ScienceOn
8 O'Carra, P. and S. Barry. 1972. Affinity chromatography of lactate dehydrogenase: model studies demonstrating the potential of the technique in the mechanistic investigation as well as in the purification of multi-substrate enzymes.FEBS Letters 21, 281-285   DOI   ScienceOn
9 O'Carra, P., S. Barry, and E. Corcoran. 1974. Affinity chromatographic differentiation of lactate dehydrogenase isoenzymes on the basis of differential abortive complex formation. FEBS Letters 43, 163-168   DOI   ScienceOn
10 Painter, P.C., S. Van Meter, R. L. Dabbs, and G. E. Clement. 1994. Analytical evaluation and comparison of Dupont aca lactate dehydrogenase-1 (LD1) isoenzyme assay diagnostic efficiency for acute myocardial infarction detection with other LD1 methods and aca CK-MB. A two-site study. Angiology 45, 585-595   DOI   ScienceOn
11 Park, H. Y. et al. 1995. Fish Biology. pp. 478, Jungmunkag Co., Korea
12 Park, S. Y. and J. J. Yum. 1993. Lactate dehydrogenase isozymes of Cypriniform and Perciform fishes: Expression of the Ldh-C gene. J. Ind. Sci. Cheongju Univ. Korea 11, 265-277
13 Park, S. Y. and J. J. Yum. 1995. Acclimation of lactate dehydrogenase isozymes in Coreoperca herzi by environmental variation. Korean J. Environ. Biol. 13, 121-130
14 Pesce, A., T. P. Fondy, F. Stolzenbach, F. Castillo and N. O. Kaplan. 1967. The comparative enzymology of lactic dehydrogenases. III. Properties of the H4 and M4 enzymes from a number of vertebrates. J. Biol. Chem. 242, 2151-2167   DOI   ScienceOn
15 Pesce, A., R. H. Mckay, F. Stolzenbach, R. D. Cahn, and N. O. Kaplan. 1964. The comparative enzymology of lactic dehydrogenases. I. Properties of the crystalline beef and chicken enzymes. J. Biol. Chem. 239, 1753-1761
16 Kim, J.-B., S. K. Cho, and J. J. Yum. 2004. Changes of activities and isozymes of lactate dehydrogenase in Coreoperca herzi to acute increase of temperature for short-term period. J. Ind. Sci. Cheongju Univ. Korea 22, 43-50
17 Kim, J.-B., S. K. Kim, and J. J. Yum. 2003. Changes of activities and isozymes of lactate dehydrogenase in Pseudogobio esocinus acclimated to acute change of temperature. J. Ind. Sci., Cheongju Univ. Korea 21, 37-44
18 Kim, M. O. and J. J. Yum. 1989. Purification, kinetics and immunochemistry of two homotetrameric lactate dehydrogenase isozymes in Pseudogobio esocinus (Cypriniformes). Korean J. Zool. 32, 420-428
19 Kolev, Y., H. Uetake, Y. Takagi, and K. Sugihara. 2008. Lactate dehydrogenase-5 (LDH-5) expression in human gastric cancer: association with hypoxia-inducible factor (HIF-1alpha) pathway, angiogenic factors production and poor prognosis. Ann. Surg. Oncol. 15, 2336-2344   DOI
20 Koukourakis, M. I., A. Giatromanolaki, C. Simopoulos, A. Polychronidis, and E. Sivridis. 2005. Lactate dehydrogenase 5 (LDH5) relates to up-regulated hypoxia inducible factor pathway and metastasis in colorectal cancer. Clin. Exp. Metastasis 22, 25-30   DOI   ScienceOn
21 Koukourakis, M. I., M. Pitiakoudis, A. Giatromanolaki, A. Tsarouha, A. Polychronidis, E. Sivridis, and C. Simopoulos. 2006. Oxygen and glucose consumption in gastrointestinal adenocarcinomas: correlation with markers of hypoxia, acidity and anaerobic glycolysis. Cancer Sci. 97, 1056-1060   DOI   ScienceOn
22 Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685   DOI   ScienceOn
23 Lam, C. W., M. H. Chan, and C. K. Wong. 2004. Severe acute respiratory syndrome: clinical and laboratory manifestations. Clin. Biochem. Rev. 25, 121-132
24 Fantin, V. R., J. St-Pierre, and P. Leder. 2006. Attenuation of LDH-A expression uncovers a link between glycolysis, mitochondrial physiology, and tumor maintenance. Cancer Cell 9, 425-434   DOI   ScienceOn
25 Fields, P. A. 2001. Protein function at thermal extremes: balancing stability and flexibility. Comp. Biochem. Physiol. A 129, 417-431   DOI   ScienceOn
26 Fields, P. A. and D. E. Houseman. 2004. Decreases in activation energy and substrate affinity in cold-adapted A4-lactate dehydrogenase: evidence from the Antarctic notothenioid fish Chaenocephalus aceratus. Mol. Biol. Evol. 21, 2246-2255   DOI   ScienceOn
27 Fields, P. A. and G. N. Somero. 1998. Hot spots in cold adaptation: localized increases in conformational flexibility in lactate dehydrogenase A4 orthologs of Antarctic notothenioid fishes. Proc. Natl. Acad. Sci. USA 95, 11476-11481   DOI   ScienceOn
28 Hochachka, P. W. and G. N. Somero. 2002. Biochemical adaptation. pp. 466, Oxford Univ. Press Inc., New York
29 Holbrook, J. J., A. Liljas, S. J. Steindel, and M. G. Rossmann. 1975. Lactate Dehydrogenase, pp. 191-192, In Boyer P. D. (ed.), The Enzymes, 3rd eds., Vol. XI, Academic Press Inc., New York
30 Holland, L. Z., M. McFall-Ngai, and G. N. Somero. 1997. Evolution of lactate dehydrogenase-A homologs of barracuda fishes (genus Sphyraena) from different thermal environments: differences in kinetic properties and thermal stability are due to amino acid substitutions outside the active site. Biochemistry 36, 3207-3215   DOI   ScienceOn
31 Johns, G. C. and G. N. Somero. 2004. Evolutionary convergence in adaptation of proteins to temperature: A4-lactate dehydrogenases of Pacific damselfishes (Chromis spp.). Mol. Biol. Evol. 21, 314-320   DOI   ScienceOn
32 Bouafia, F., J. Drai, J. Bienvenu, C. Thieblemont, D. Espinouse, G. Salles, and B. Coiffier. 2004. Profiles and prognostic values of serum LDH isoenzymes in patients with haematopoietic malignancies. Bull. Cancer 91, E229-240
33 Bradford, M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248-254   DOI   ScienceOn
34 Cho, S. K. 2000. Mitochondrial lactate dehydrogenase in tissues of vertebrate. 88pp., Ph.D. Thesis Cheongju Univ., Korea.
35 Cho, S. K., S. Y. Park, and J. J. Yum. 1993. Purification and immunochemistry of lactate dehydrogenase isozyme in Lampetra japonica. Korean J. Zool. 36, 505-513
36 Cho, S. K. and J. J. Yum. 1993. Heterogeneity of lactate dehydrogenase isozymes in tissues of Lampetra japonica. Korean J. Zool. 36, 319-328
37 Cho, S. K. and J. J. Yum. 1996. The adaptational phenotype of lactate dehydrogenase isozymes in Pseudogobio esocinus by the environmental variation. J. Ind. Sci., Cheongju Univ. Korea 14, 333-343
38 Cho, S. K. and J. J. Yum. 2004. Lactate dehydrogenase isozyme of hypoxia tropical catfish (Pangasius polyuranodon, Hypostomus plecostomus). J. Life Sci. 14, 702-707   DOI
39 Cho, S. K. and J. J. Yum. 2005. Changes of activities and isozymes of lactate dehydrogenase in Coreoperca herzi and Pseudogobio esocinus acclimated to rapid increase of dissolved oxygen. J. Life Sci. 15, 71-79   DOI
40 Davis, B. J. 1964. Disc electrophoresis-Ⅱ. Method and application to human serum proteins. Ann. N.Y. Acad. Sci. 121, 404-427   DOI
41 Donovan, J. and P. Brown. 1991. Cardiac Puncture of Rabbit, Unit 1.7.6., In Coligan, J. E. et al. (eds.), Current Protocols in Immunology, Vol. 1, John Wiley & Sons Inc., New York.
42 Almeida-Val, V. M. F. and A. L. Val. 1993. Evolutionary trends of LDH isozymes in fishes. Comp. Biochem. Physiol. B 105, 21-28   DOI
43 Andriutsa, K. A. and A. K. Andriutsa. 1987. Diagnostic and pathogenetic significance of determining lactate de hydrogenase isoenzyme activity in various biological media in patients with viral hepatitis. Ter. Arkh. 59, 89-94