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http://dx.doi.org/10.5352/JLS.2011.21.11.1565

Purification and Characterization of Eye-Specific Lactate Dehydrogenase C4 Isozyme in Greenling (Hexagrammos otakii)  

Cho, Sung-Kyu (Industrial Science Research Institute, Cheongju University)
Yum, Jung-Joo (Department of Life Science, Cheongju University)
Publication Information
Journal of Life Science / v.21, no.11, 2011 , pp. 1565-1572 More about this Journal
Abstract
Eye-specific lactate dehydrogenase (EC 1.1.1.27, LDH) $C_4$ isozyme in the eyes of greenlings (Hexagrammos otakii) was successfully purified by affinity chromatography and continuous-elution electrophoresis. The molecular weight of the purified eye-specific LDH $C_4$ isozyme was 154.8 kDa, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Optimal pH for enzymatic reaction of the eye-specific LDH $C_4$ isozyme was pH 8.5. $K^{PYR}_m$ value of the purified eye-specific LDH $C_4$ isozyme was $1.88{\times}10^{-5}$ M using pyruvate as a substrate. These results indicate that we must consider pH when measuring eye-specific LDH $C_4$ isozyme activity. The eye-specific LDH $C_4$ isozyme had a higher binding affinity for the substrate as a pyruvate than LDH A4 isozyme. Antibodies produced against the purified eye-specific LDH $C_4$ isozyme may be used in the diagnosis of several human diseases and in comparative physiological studies of fishes.
Keywords
Greenling (Hexagrammos otakii); lactate dehydrogenase (LDH); eye-specific LDH $C_4$ isozyme;
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1 Whitt, G. S. 1970. Developmental genetics of the lactate dehydrogenase isozymes of fish. J. Exp. Zool. 175, 1-35.   DOI   ScienceOn
2 Whitt, G. S. 1981. Evolution of isozyme loci and their differential regulation, pp. 271-289, In Scudder, G. G. E. and J. L. Reveal (eds.), Evolution today: proceedings of second international congress of systematic and evolutionary biology. Hunt Institute for Botanical Documentation, Carnegie-Mellon Univ. Press, Pittsburgh.
3 Park, S. Y., S. K. Cho, and J. J. Yum. 2004. Characterization and evolutionary relationship of lactate dehydrogenase in liver of Lampetra japonica and liver-specific $C_{4}$ isozyme in Gadus macrocephalus. J. Life Sci. 14, 708-715.   DOI
4 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.
5 Park, S. Y. and J. J. Yum. 1997. Purification and characterization of lactate dehydrogenase eye- and testis-specific $C_{4}$ isozyme. J. Ind. Sci., Cheongju Univ., Korea. 15, 263-270.
6 Park, E. M. and J. J. Yum. 2010. Purification and characterization of lactate dehydrogenase isozymes in Channa argus. J. Life Sci. 20, 260-268.   DOI
7 Park, E. M. and J. J. Yum. 2011. Activities of lactate dehydrogenase and ratios of lactate dehydrogenase/citrate synthase in tissues of Odontobutis interrupta. J. Ind. Sci., Cheongju Univ., Korea. 28, 15-24.
8 Rehse, P. H. and W. S. Davidson. 1986. Purification and properties of a C-type isozyme of lactate dehydrogenase from the liver of the Atlantic cod (Gadus morhua). Comp. Biochem. Physiol. B 84, 145-150.   DOI
9 Sensabaugh, G. F. and N. O. Kaplan. 1972. A lactate dehydeogenase specific to the liver of gadoid fish. J. Biol. Chem. 247, 585-593.
10 Shaklee, J. B., K. L. Kepes, and G. S. Whitt. 1973. Specialized lactate dehydrogenase isozymes: the molecular and genetic basis for the unique eye and liver LDHs of teleost fishs. J. Exp. Zool. 185, 217-240.   DOI   ScienceOn
11 Kim, J. B., S. K. Cho, and J. J. Yum. 2004. Changes of activities and isozymes of lactate dehydrogenase in Coreoperca herzi acclimated to acute increase of temperature for short-term period. J. Ind. Sci., Cheongju Univ., Korea. 22, 43-50.
12 Davis, B. J. 1964. Disc electrophoresis-II. Method and application to human serum proteins. Ann. N.Y. Acad. Sci. 121, 404-427.   DOI
13 Coonrod, S., A. Vitale, C. Duan, S. Bristol-Gould, J. Herr, and E. Goldberg. 2006. Testis-specific lactate dehydrogenase (LDH-$C_{4}$; Ldh3) in murine oocytes and preimplantation embryos. J. Androl. 27, 502-509.   DOI   ScienceOn
14 Coppes, Z. 1992. Lactate dehydrogenase in teleosts: the role of LDH-$C_{4}$ isozyme. Comp. Biochem. Physiol. B 102, 673-677.   DOI
15 Coronel, C. E., C. Burgos, N. M. Gerez de Burgos, L. E. Rovai, and A. Blanco. 1983. Catalytic properties of the sperm-specific lactate dehydrogenase (LDH X or $C_{4}$) from different species. J. Exp. Zool. 225, 379-385.   DOI   ScienceOn
16 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.
17 Fisher, S. E., J. B. Shaklee, S. D. Ferris, and G. S. Whitt. 1980. Evolution of five multilocus isozyme systems in the chordates. Genetica 52/53, 73-85.   DOI
18 Goldberg, E. 1972. Amino acid composition and properties of crystalline lactate dehydrogenase X from mouse testes. J. Biol. Chem. 247, 2044-2048.
19 Goldberg, E., E. M. Eddy, C. Duan, and F. Odet. 2010. LDHC: the ultimate testis-specific gene. J. Androl. 31, 86-94.   DOI   ScienceOn
20 Higuti, I. H., S. W. Grande, R. Sacco, and A. J. Nascimento. 2003. Isolation of alkalophilic CGTase-producing bacteria and characterization of cyclodextrin-glycosyltransferase. Braz. Arch. Biol. Technol. 46, 183-186.   DOI   ScienceOn
21 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.
22 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
23 Andriutsa, K. A. and A. K. Andriutsa. 1987. Diagnostic and pathogenetic significance of determining lactate dehydrogenase isoenzyme activity in various biological media in patients with viral hepatitis. Ter. Arkh. 59, 89-94.
24 Baldwin, J. and P. S. Lake. 1987. Lactate dehydrogenase homopolymer of hagfish heart and the single lactate dehydrogenase of lampreys display greater immunochemical similarity to LDH $C_{4}$ than to LDH $B_{4}$ of teleost fish. J. Exp. Zool. 242, 99-102.   DOI
25 Baldwin, J., K. Mortimer, and A. Patak. 1988. Do ascidians possess the ancestral subunit type of vertebrate lactate dehydrogenase. J. Exp. Zool. 246, 109-114.   DOI
26 Cho, S. K. 2000. Mitochondrial lactate dehydrogenase in tissues of vertebrate. Ph. D. Thesis, Cheongju University, Cheongju, Korea.
27 Cho, S. K., B. Ku, H. An, E. M. Park, S. Y. Park, J. B. Kim, and J. J. Yum. 2009. Purification and characterization of lactate dehydrogenase $A_{4}$ isozyme in mandrin fish (Siniperca scherzeri). J. Life Sci. 19, 256-263.   DOI
28 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.
29 Cho, S. K. and J. J. Yum. 1993. Heterogeneity of lactate dehydrogenase isozymes in tissues of Lampetra japonica. Korean J. Zool. 36, 319-328.
30 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
31 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.
32 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
33 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
34 An, C. S., S. K. Cho, and J. J. Yum. 2010. Variation of lactate dehydrogenase isozymes in angelfish (Pterophyllum scalare) according to acute environmental change. J. Life Sci. 20, 416-423.   DOI
35 Yum, J. J. 2008. Characterization of lactate dehydrogenase in Acanthogobius hasta. J. Life Sci. 18, 264-272.   DOI
36 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
37 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.
38 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-1$\alpha$) pathway, angiogenic factors production and poor prognosis. Ann. Surg. Oncol. 15, 2336-2344.   DOI   ScienceOn
39 Koslowski, M, O. Tureci, C. Bell, P. Krause, H. A. Lehr, J. Brunner, G. Seitz, F. O. Nestle, C. Huber, and U. Sahin. 2002. Multiple splice variants of lactate dehydrogenase C selectively expressed in human cancer. Cancer Res. 62, 6750-6755.
40 Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage $T_{4}$. Nature 227, 680-685.   DOI   ScienceOn
41 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
42 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
43 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