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Connexin 36 in bovine retina: Lack of phosphorylation but evidence for association with phosphorylated proteins

Published online by Cambridge University Press:  18 November 2003

ARI SITARAMAYYA ,
JOHN W. CRABB ,
DIANE F. MATESIC ,
ALEXANDER MARGULIS ,
VINITA SINGH ,
SADHONA PULUKURI  and
LOAN DANG
ARI SITARAMAYYA
Affiliation:
Eye Research Institute, Oakland University, Rochester
JOHN W. CRABB
Affiliation:
Cole Eye Institute, Cleveland Clinic Foundation, Cleveland
DIANE F. MATESIC
Affiliation:
Department of Pharmaceutical Sciences, Mercer University, Atlanta
ALEXANDER MARGULIS
Affiliation:
Eye Research Institute, Oakland University, Rochester
VINITA SINGH
Affiliation:
Eye Research Institute, Oakland University, Rochester
SADHONA PULUKURI
Affiliation:
Eye Research Institute, Oakland University, Rochester
LOAN DANG
Affiliation:
Eye Research Institute, Oakland University, Rochester
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Abstract

In vertebrate retina interneuronal communication through gap junctions is involved in light adaptation and in the transfer of visual information from the rod pathway to the cone pathway. Reports over the last two decades have indicated that these gap junctions are regulated by cyclic nucleotide-dependent protein kinases suggesting that the gap junction proteins, connexins, are phosphorylated. Though all the connexins involved in light adaptation and information transfer from rod to cone pathway are not yet known, connexin 36 has been shown to be definitively involved in the latter process. We have therefore attempted to investigate the cyclic nucleotide-dependent phosphorylation of this connexin in bovine retina. We found several soluble and membrane proteins in bovine retina whose phosphorylation was regulated by cyclic nucleotides. However, no protein of about 36 kDa with cyclic nucleotide-regulated phosphorylation was found in gap junction-enriched membrane preparations. A 36-kDa phosphorylated protein was found in gap junction-enriched membranes phosphorylated in the presence of calcium. However, this protein was not immunoprecipitated by anti-connexin 36 antibodies indicating that it was not connexin 36 in spite of its similarity in molecular weight. Immunoprecipitation did reveal phosphorylated proteins coimmunoprecipitated with connexin 36. Two of these proteins were identified as beta and alpha tubulin subunits. Though cyclic GMP and calcium did not greatly influence the association of these proteins with connexin 36, the results suggest the possibility of connexin 36 associating with other proteins. Together, these observations indicate that interneuronal communication at gap junctions made by connexin 36 may not be regulated by direct phosphorylation of connexin 36, but possibly by phosphorylation of associated proteins.

Keywords

Connexin 36Gap junctionsConnexin-associated proteinsPhosphorylationRetina
Type
Research Article
Information
Visual Neuroscience , Volume 20 , Issue 4 , July 2003 , pp. 385 - 395
Copyright
2003 Cambridge University Press

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