Abstract
Photopic (cone) vision essential for color sensation, central vision, and visual acuity is mediated by the activation of photoreceptor cyclic nucleotide-gated (CNG) channels. Naturally occurring mutations in the cone channel subunits CNGA3 and CNGB3 are associated with achromatopsia and cone dystrophies. This work investigated the functional modulation of cone CNG channel by exploring the channel-interacting proteins. Retinal protein extracts prepared from cone-dominant Nrl −/− mice were used in CNGA3 antibody affinity purification, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) separation and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry analysis. The peptide mass fingerprinting of the tryptic digests and database search identified a number of proteins including spectrin alpha-2, ATPase (Na+/K+ transporting) alpha-3, alpha and beta subunits of ATP synthase (H+ transporting, mitochondrial F1 complex), and alpha-2 subunit of the guanine nucleotide-binding protein. In addition, the affinity-binding assays demonstrated an interaction between cone CNG channel and calmodulin but not cone Na+/Ca2+–K+ exchanger in the mouse retina. Results of this study provide insight into our understanding of cone CNG channel-interacting proteins and the functional modulations.
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Acknowledgments
This work was supported by grants from the National Center for Research Resources (P20RR017703), the National Eye Institute (EY12190, EY019490, and EY17888), and the Oklahoma Center for the Advancement of Science & Technology (OCAST). We thank Drs. Anand Swaroop and Jonathan Lytton for providing Nrl −/− mouse line and anti-NCKX2 antibody, respectively.
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Ding, XQ., Matveev, A., Singh, A., Komori, N., Matsumoto, H. (2014). Exploration of Cone Cyclic Nucleotide-Gated Channel-Interacting Proteins Using Affinity Purification and Mass Spectrometry. In: Ash, J., Grimm, C., Hollyfield, J., Anderson, R., LaVail, M., Bowes Rickman, C. (eds) Retinal Degenerative Diseases. Advances in Experimental Medicine and Biology, vol 801. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-3209-8_8
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DOI: https://doi.org/10.1007/978-1-4614-3209-8_8
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