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. 1990 Sep;94(1):109–113. doi: 10.1104/pp.94.1.109

Immunocharacterization of NADH-Glutamate Dehydrogenase from Vitis vinifera L

C A Loulakakis 1,2, K A Roubelakis-Angelakis 1,2
PMCID: PMC1077197  PMID: 16667675

Abstract

Rabbit antiserum was raised against NADH-glutamate dehydrogenase (GDH) isoenzyme 1, purified from leaves of Vitis vinifera L. cv Soultanina and its specificity was tested. This antiserum was used for immunocharacterization of the GDH from leaf, shoot, and root tissues. The antiserum recognized the seven isoenzymes of NADH-GDH and precipitated all the enzyme activity from the three tissues tested. Western blot following SDS-PAGE revealed the same protein band for the three tissues, with a molecular mass of 42.5 kilodaltons corresponding to NADH-GDH subunit. Results, based on the immunological studies, revealed that NADH-GDH from leaf, shoot, and root tissues are closely related proteins. Furthermore, addition of ammonium ions to the culture medium of in vitro grown explants resulted in a significant increase in NADH-GDH activity in root, shoot, and leaf tissues.

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Selected References

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  1. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  2. Loyola-Vargas V. M., de Jimenez E. S. Differential role of glutamate dehydrogenase in nitrogen metabolism of maize tissues. Plant Physiol. 1984 Oct;76(2):536–540. doi: 10.1104/pp.76.2.536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Nagel M., Hartmann T. Glutamate dehydrogenase from Medicago sativa L., purification and comparative kinetic studies of the organ-specific multiple forms. Z Naturforsch C. 1980 May-Jun;35(5-6):406–415. doi: 10.1515/znc-1980-5-610. [DOI] [PubMed] [Google Scholar]
  4. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Yamaya T., Oaks A., Rhodes D., Matsumoto H. Synthesis of [N]glutamate from [N]h(4) and [N]glycine by mitochondria isolated from pea and corn shoots. Plant Physiol. 1986 Jul;81(3):754–757. doi: 10.1104/pp.81.3.754. [DOI] [PMC free article] [PubMed] [Google Scholar]

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