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Z. Li, H. Lu, G. Su, Y. Du, P. Yu; Expressions and Protein Interaction Between Neuroglobin and Na+, K+-ATPase Beta 2 Subunit in Rat Retina. Invest. Ophthalmol. Vis. Sci. 2008;49(13):4940.
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© ARVO (1962-2015); The Authors (2016-present)
Neuroglobin is a newly discovered nervous system specific O2-binding protein, and it’s concentration in the retina is 100-fold higher than that in total brain extracts. Na+, K+-ATPase beta 2 subunit was identified to have interaction with neuroglobin by east two hybrid technique in vitro. This study was designed to investigate the expressions and protein interaction between neuroglobin and Na+, K+-ATPase beta 2 subunit in vivo in rat retinas.
The polyclonal antibody against Na+, K+-ATPase beta 2 subunit and anti-neuroglobin monoclonal antibodies were produced using Japanese adult male rabbits and immunizing Balb/c mice respectively. We detected the expression of the above two proteins by gel electrophoresis and western blot skills and the interaction was proved by co-immunoprecipitation test in vivo.
The homemade antiserums could detect both of the neuroglobin and Na+, K+-ATPase beta 2 subunit proteins which expressed in rat retinas. The apparent molecular mass of native neuroglobin in retina is 17 kDa and for Na+, K+-ATPase beta 2 subunit, it is 45kDa. Immunoprecipitation product was detected by using Western blot with either anti-neuroglobin monoclonal antibody or polyclonal antibody against Na+, K+-ATPase beta 2 subunit.
Neuroglobin, an oxygen-binding respiratory protein, is synthesized at a high level in the rat retina and there is an interaction between the former protein and the β2 subunit of Na+-K+-ATPase, of which both might attend the activities of oxygen supply or consumption and maintain the ion homeostasis in retinas.
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