May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Molecular Analysis of a Consensus Sequence for Casein Kinase 2 (CK2) in the NR1 Subunit of Retina NMDA Receptor
Author Affiliations & Notes
  • A. Lopez-Colome
    Neuroscience, Instituto de Fisiologia Celular, Universidad Nacional Autónoma de México, Mexico City, Mexico
  • A. Parrales
    Neuroscience, Instituto de Fisiologia Celular, Universidad Nacional Autónoma de México, Mexico City, Mexico
  • Footnotes
    Commercial Relationships  A. Lopez-Colome, None; A. Parrales, None.
  • Footnotes
    Support  Partially supported by Grants IN228203 (PAPIIT) and 42640-Q (CONACyT) to A.M.L.C.
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 3167. doi:
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    • Get Citation

      A. Lopez-Colome, A. Parrales; Molecular Analysis of a Consensus Sequence for Casein Kinase 2 (CK2) in the NR1 Subunit of Retina NMDA Receptor. Invest. Ophthalmol. Vis. Sci. 2008;49(13):3167.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : Glutamate is the major excitatory neurotransmitter in the vertebrate retina and exerts its effects through the activation of distinct receptor subtypes located on postsynaptic neurons. The calcium-permeable NMDAR subtype plays a central role in retinal transmission, and has been implicated in differentiation, plasticity and excitotoxicity in this tissue. We have identified a novel alternatively spliced cassette (C3) in the C-terminal region of the channel-forming NR1 subunit of the NMDAR in chick retina, coding for a consensus sequence for CK2 phosphorylation (EETSEH). Importantly, CK2 has been implicated in cell cycle and circadian rhythms, in which the retina plays a key role. The purpose of this study was to develop a model system for the in vitro analysis of the NR1subunit C3 cassette phosphorylation by CK2 in the retina.

Methods: : We generated fusion proteins of NR1 C-terminal containing or lacking the C3 cassette, with Xpress epitope. These constructs were overexpressed in HEK cells, and CK2 phosphorylation was analyzed for both proteins. Finally, co-immunoprecipitation of CK2 with the fusion proteins was probed.

Results: : Using this model, we demonstrated that CK2 and C3 cassette of NR1 subunit do not interact, in spite of the presence of the consensus sequence.

Conclusions: : We conclude that such failure to interact is due to the presence of a basic residue (His) within the C3 cassette, and of a stop codon following CK2 consensus sequence. These results suggest that proteins bearing a consensus sequence for CK2 phosphorylation require the presence of acidic residues at the C-terminal of the target sequence in order to achieve functional activation.

Keywords: receptors • excitatory neurotransmitters • protein structure/function 
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