Detection of active proteasome structures in brain extracts: proteasome features of August rat brain with violations in monoamine metabolism
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Pavel A. Erokhov1,*, Yulia V. Lyupina1,*, Alexandra S. Radchenko1, Anna A. Kolacheva2, Yulia O. Nikishina2 and Natalia P. Sharova1
1Laboratory of Biochemistry of Ontogenesis Processes, N.K. Koltsov Institute of Developmental Biology of Russian Academy of Sciences, Moscow, Russia
2Laboratory of Neural and Neuroendocrine Regulations, N.K. Koltsov Institute of Developmental Biology of Russian Academy of Sciences, Moscow, Russia
*These authors have contributed equally to this work
Natalia P. Sharova, email: [email protected]
Keywords: proteasome structure, immune proteasomes, rat brain, violations in monoamine metabolism, native electrophoresis
Received: May 26, 2017 Accepted: July 23, 2017 Published: August 10, 2017
The aim of this work was to detect changes in proteasome pools of brain parts of August rats with monoamine metabolism violations in comparison with that of control Wistar rats. To reveal active proteasome structures, a method of native electrophoresis for the analysis of crude tissue fractions was developed. By means of this method and following Western blotting, the most pronounced changes in reorganization of proteasome structures were detected in proteasome pool of the brain cortex of August rats. Main findings are the enhanced expression of immune proteasome subtypes containing proteolytic subunit LMP2 and activator PA28αβ as well as immune proteasome subtypes containing proteolytic subunit LMP7 and activator PA700 and simultaneously decreased expression of subtypes with subunit LMP2 and activator PA700 in the brain cortex of August rats compared to that of Wistar rats. These results were indirectly confirmed by SDS PAGE method followed by Western blotting, which showed the increased quantities of immune subunits and proteasome activators in the brain cortex of August rats compared to that of Wistar rats. Immune proteasomes were revealed by immunohistochemistry in neurons, but not in glial cells of August and Wistar rat cortex. The detected reorganization of proteasome pools is likely to be important for production of special peptides to provide the steady interaction between neurons and adaptation of central nervous system to conditions caused by monoamine metabolism deviations.
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