Characterization of Nuclear Glutamate Dehydrogenase of Chicken Liver and Brain

Pragnya      Panda; Mrutyunjay      Suar; Divya      Singh; Shashibhal      Matacharan Pandey; Madan      Mohan Chaturvedi; Jogeswar      Satchidananda Purohit

Abstract

Glutamate dehydrogenase (GDH) enzyme is recently being reported to be present in the nucleus in addition to the mitochondria in a number of organisms. Here we investigated the distribution of GDH in liver and brain tissues of chicken. Polyclonal anti-GDH antibody against bovine GDH was raised by us, which was later shown to be immunereactive to chicken GDH. The nuclear and the mitochondrial extracts from liver and brain tissues of chicken were made as described. By quantitative immunoreactivity, it was revealed that the nuclear GDH expressed in comparable efficiencies in the liver and brain. However, the activity of the brain nuclear GDH was lower than the liver counterparts. The allosteric regulation pattern for the brain nuclear GDH was also different from the other corresponding fractions and it was speculated that the brain nuclear GDH was inactive. The liver and brain nuclear GDH were purified to homogeneity and comparison of specific activities of both the GDH ruled out the existence of any inhibitor in the brain nuclear GDH. It is hypothesized that the inactivation of the brain nuclear GDH in chicken could be due to some already known posttranslational modification. The present report throws light on the differential regulation pattern of GDH enzyme.

Keywords: Glutamate dehydrogenase (GDH), GDH inactivation, immunoreactivity, NAD(H), GTP, cAMP-dependent, ADP ribosyl transferase, nuclear and mitochondrial extractsQ, SDS-PAGE, Electrophoresis, Miyazaki, GLUD1, RT PCR, NADH controls, CIAPGlutamate dehydrogenase (GDH), GDH inactivation, immunoreactivity, NAD(H), GTP, cAMP-dependent, ADP ribosyl transferase, nuclear and mitochondrial extractsQ, SDS-PAGE, Electrophoresis, Miyazaki, GLUD1, RT PCR, NADH controls, CIAP