INVOLVEMENT OF THE RECOVERIN C-TERMINAL SEGMENT IN RECOGNITION OF THE TARGET ENZYME RHODOPSIN KINASE

Zernii, E.Y.; Komolov, K.E.; Permyakov, S.E.; Kolpakova, T.; Dell'Orco, D.; Poetzsch, A.; Knyazeva, E.L.; Grigoriev, I.I.; Permyakov, E.A.; Senin, I.I.; Philippov, P.P.; Koch, K.W.

Авторы: Zernii E.Y., Komolov K.E., Permyakov S.E., Kolpakova T., Dell'Orco D., Poetzsch A., Knyazeva E.L., Grigoriev I.I., Permyakov E.A., Senin I.I., Philippov P.P., Koch K.W.
Журнал: Biochemical Journal
Том: 435
Номер: 2
Год издания: 2011
Издательство: Portland Press, Ltd.
Местоположение издательства: United Kingdom
Первая страница: 441
Последняя страница: 450
DOI: 10.1042/BJ20110013
Аннотация: NCS (neuronal Ca2+ sensor) proteins belong to a family of calmodulin-related EF-hand Ca2+-binding proteins which, in spite of a high degree of structural similarity, are able to selectively recognize and regulate individual effector enzymes in a Ca2+-dependent manner. NCS proteins vary at their C-termini, which could therefore serve as structural control elements providing specific functions such as target recognition or Ca2+ sensitivity. Recoverin, an NCS protein operating in vision, regulates the activity of rhodopsin kinase, GRK1, in a Ca2+-dependent manner. In the present study, we investigated a series of recoverin forms that were mutated at the C-terminus. Using pull-down assays, surface plasmon resonance spectroscopy and rhodopsin phosphorylation assays, we demonstrated that truncation of recoverin at the C-terminus significantly reduced the affinity of recoverin for rhodopsin kinase. Site-directed mutagenesis of single amino acids in combination with structural analysis and computational modelling of the recoverin-kinase complex provided insight into the protein-protein interface between the kinase and the C-terminus of recoverin. Based on these results we suggest that Phe3 from the N-terminal helix of rhodopsin kinase and Lys192 from the C-terminal segment of recoverin form a cation-π interaction pair which is essential for target recognition by recoverin. Taken together, the results of the present study reveal a novel rhodopsin-kinase-binding site within the C-terminal region of recoverin, and highlights its significance for target recognition and regulation.
Добавил в систему: Сенин Иван Иванович

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INVOLVEMENT OF THE RECOVERIN C-TERMINAL SEGMENT IN RECOGNITION OF THE TARGET ENZYME RHODOPSIN KINASEстатья

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