Allosteric Communication of the Dimerization and the Catalytic Domain in Photoreceptor Guanylate Cyclase.

Phototransduction in vertebrate photoreceptor cells is controlled by Ca-dependent feedback loops involving the membrane-bound guanylate cyclase GC-E that synthesizes the second messenger guanosine-3',5'-cyclic monophosphate. Intracellular Ca-sensor proteins named guanylate cyclase-activating proteins (GCAPs) regulate the activity of GC-E by switching from a Ca-bound inhibiting state to a Ca-free/Mg-bound activating state. The gene encodes for human GC-E, and mutations in are often associated with an imbalance of Ca and cGMP homeostasis causing retinal disorders. Here, we investigate the Ca-dependent inhibition of the constitutively active GC-E mutant V902L. The inhibition is not mediated by GCAP variants but by Ca replacing Mg in the catalytic center. Distant from the cyclase catalytic domain is an α-helical domain containing a highly conserved helix-turn-helix motif. Mutating the critical amino acid position 804 from leucine to proline left the principal activation mechanism intact but resulted in a lower level of catalytic efficiency. Our experimental analysis of amino acid positions in two distant GC-E domains implied an allosteric communication pathway connecting the α-helical and the cyclase catalytic domains. A computational connectivity analysis unveiled critical differences between wildtype GC-E and the mutant V902L in the allosteric network of critical amino acid positions.