NADP(+)-activable, NAD(+)-specific glutamate dehydrogenase. Purification and immunological analysis.
作者： B. Yang , H.B. LéJohn
DOI: 10.1016/S0021-9258(17)41807-2
关键词: Enzyme activator 、 Oxidative deamination 、 Achlya klebsiana 、 Biology 、 NAD+ kinase 、 Molecular biology 、 Sepharose 、 Biochemistry 、 Enzyme 、 Protein subunit 、 Glutamate dehydrogenase
摘要: An NAD(+)-specific glutamate dehydrogenase (NAD-GDH) that is inducible by L-glutamine was isolated from Achlya klebsiana and purified to electrophoretic homogeneity. The enzyme only partially active in vitro unless NADP+ (an activator) present both its oxidative deamination reductive amination reactions. This type of reported (LeJohn, H.B. (1971) Nature 231, 164-168) be widespread among the amorphous group algae-related fungi classified as Oomycota. retained dependence on at all stages purification. decreased Km substrates 3-fold increased Vmax 4-fold. M(r) undernatured 480,000, and, denatured, a single subunit 120,000 seen. A polyclonal antibody raised rabbit against excised SDS-polyacrylamide gel electrophoresis gels immunoprecipitated polypeptide, undenatured enzyme, physically distinct polypeptide 74,000. antibody, anchored antigen Sepharose, still 74,000 polypeptide. found 220,000 native protein.
sci-hub.st 参考文章(21)
Beatrice Dewald, John T. Dulaney, Oscar Touster, Solubilization and polyacrylamide gel electrophoresis of membrane enzymes with detergents. Methods in Enzymology. ,vol. 32, pp. 82- 91 ,(1974) , 10.1016/0076-6879(74)32011-3
Roselynn M. Stevenson, H.B. LéJohn, Glutamic Dehydrogenases of Oomycetes KINETIC MECHANISM AND POSSIBLE EVOLUTIONARY HISTORY Journal of Biological Chemistry. ,vol. 246, pp. 2127- 2135 ,(1971) , 10.1016/S0021-9258(19)77198-1
H.B. LéJohn, L.E. Cameron, B. Yang, S.L. Rennie, Molecular characterization of an NAD-specific glutamate dehydrogenase gene inducible by L-glutamine. Antisense gene pair arrangement with L-glutamine-inducible heat shock 70-like protein gene. Journal of Biological Chemistry. ,vol. 269, pp. 4523- 4531 ,(1994) , 10.1016/S0021-9258(17)41809-6
Cleland Ww, The kinetics of enzyme-catalyzed reactions with two or more substrates or products. I. Nomenclature and rate equations. Biochimica et Biophysica Acta. ,vol. 67, pp. 104- 137 ,(1963) , 10.1016/0006-3002(63)91800-6
H.B. LéJohn, L.E. Cameron, B. Yang, G. MacBeath, D.S. Barker, S.A. Williams, Cloning and analysis of a constitutive heat shock (cognate) protein 70 gene inducible by L-glutamine. Journal of Biological Chemistry. ,vol. 269, pp. 4513- 4522 ,(1994) , 10.1016/S0021-9258(17)41808-4
Richard D. Palmiter, Takami Oka, Robert T. Schimke, Modulation of Ovalbumin Synthesis by Estradiol-17β and Actinomycin D as Studied in Explants of Chick Oviduct in Culture Journal of Biological Chemistry. ,vol. 246, pp. 724- 737 ,(1971) , 10.1016/S0021-9258(18)62473-1
H.B. LéJohn, Roselynn M. Stevenson, Multiple Regulatory Processes in Nicotinamide Adenine Dinucleotide-specific Glutamic Dehydrogenases Journal of Biological Chemistry. ,vol. 245, pp. 3890- 3900 ,(1970) , 10.1016/S0021-9258(18)62933-3
Linda E. Cameron, H.B. LéJohn, On the Involvement of Calcium in Amino Acid Transport and Growth of the Fungus Achlya Journal of Biological Chemistry. ,vol. 247, pp. 4729- 4739 ,(1972) , 10.1016/S0021-9258(19)44973-9
Therese M. Timmons, Bonnie S. Dunbar, Protein blotting and immunodetection. Methods in Enzymology. ,vol. 182, pp. 679- 688 ,(1990) , 10.1016/0076-6879(90)82053-5
K Weber, M Osborn, The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis Journal of Biological Chemistry. ,vol. 244, pp. 4406- 4412 ,(1969) , 10.1016/S0021-9258(18)94333-4