Abstract
Human rhodopsin kinase (RK) and a carboxyl terminus-truncated mutant RK lacking the last 59 amino acids (RKC) were expressed in human embryonic kidney 293 cells to investigate the role of the carboxyl terminus of RK in recognition and phosphorylation of rhodopsin. RKC, like the wild-type RK, was detected in both plasma membranes and cytosolic fractions. The Cterminal truncated rhodopsin kinase was unable to phosphorylate photo-activated rhodopsin, but possesses kinase activity similar to the wild-type RK in phosphorylation of small peptide substrate. It suggests that the truncation did not disturb the gross structures of RK catalytic domain. Our results also show that RKC failed to translocate to photo-activated rod out segments. Taken together, our study demonstrate the carboxyl terminus of RK is required for phosphorylation of photo-activated rhodopsin and strongly indicate that carboxyl-terminus of RK may be involved in interaction with photo-activated rhodopsin.
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Abbreviations
- Rho:
-
photo-activated rhodopsin
- RK:
-
rhodopsin kinase
- ROS:
-
rod outer segments
- SDS:
-
sodium dodecyl sulfate
- PAGE:
-
polyacrylamide gel electrophoresis
- PKA:
-
cyclic adenosine monophosphate-dependent protein kinase
- PKC:
-
calcium-dependent protein kinase
- GPRK:
-
G protein-coupled receptor kinase
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Acknowledgements
This research was supported by research grants from National Natural Science Foundation of China (39630130 and 39625015), Chinese Academy of Sciences (KJ951-B1-608 and KY951-A1-301), Shanghai Research Center of Life Sciences, Shanghai Educational Development Foundation, and German Max-Planck Society. The authors wish to thank Dr. Lin Li for recombinant bovine GRK2 and Jianfang Din for technical assistance help in preparing anti-RK antibodies.
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Yu, Q., Cheng, Z., Zhao, J. et al. Carboxyl terminal of rhodopsin kinase is required for the phosphorylation of photo-activated rhodopsin. Cell Res 8, 303–310 (1998). https://doi.org/10.1038/cr.1998.30
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DOI: https://doi.org/10.1038/cr.1998.30
