title: Mechanism involved in the modulation of photoreceptor-specific cyclic nucleotidegated channel by the tyrosine kinase adapter protein Grb14.
abstract: We recently found that growth factor receptor-bound (Grb) protein 14 is a novel physiological modulator of photoreceptor specific cyclic nucleotide-gated channel alpha subunit (CNGA1). Grb14 promotes the CNG channel closure through its Ras-associating (RA) domain. In the current study we show that this RA domain-mediated inhibition of rod CNG channel is electrostatic in nature. Grb14 competes with cGMP for the CNGA1 binding pocket and electrostatically interacts with Arg(559) through a negatively charged β-turn at its RA domain. Moreover, the three Glu residues (180-182) in Grb14 are absolutely critical for electrostatic interaction with the cGMP binding pocket and resultant inhibition. Our study also demonstrates that substitution of Lys140 for Ala or in combination with polyglutamte mutants of Grb14 results in a significantly reduced binding with CNGA1. These results suggest that in addition to Glu(180-182) and Lys(140), other residues in Grb14 may be involved in the electrostatic interaction with CNGA1. The RA domain is highly conserved among the members of Grb7 family of proteins, which includes Grb7, Grb10 and Grb14. Further, only Grb14 is able to modulate the channel activity, but not Grb7 and Grb10. All together, it suggests the existence of a divergence in RA domains among the members of the Grb7 family.
authors.affiliation: Department of Ophthalmology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA.
authors.forename: Vivek K
authors.initials: VK
authors.lastname: Gupta
authors.forename: Ammaji
authors.initials: A
authors.lastname: Rajala
authors.forename: Karla K
authors.initials: KK
authors.lastname: Rodgers
authors.forename: Raju V S
authors.initials: RV
authors.lastname: Rajala
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000378:metabolism; D054815:Cyclic Nucleotide-Gated Cation Channels / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051383:GRB10 Adaptor Protein / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D008958:Models, Molecular; D011487:Protein Conformation; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D055672:Static Electricity
chemical_list: D054815:Cyclic Nucleotide-Gated Cation Channels; C096358:GRB10 protein, human; D051383:GRB10 Adaptor Protein; D011505:Protein-Tyrosine Kinases; D006152:Cyclic GMP; D002118:Calcium