Amino acid sequences of two immune-dominant epitopes of recoverin are involved in Ca2+/recoverin-dependent inhibition of phosphorylation of rhodopsin.

Antibodies AB(60-72) and AB(80-92) against two immune-dominant epitopes of photoreceptor Ca(2+)-binding protein recoverin, 60-DPKAYAQHVFRSF-72 and 80-LDFKEYVIALHMT-92, which can be exposed in a Ca(2+)-dependent manner, were obtained. The presence of AB(60-72) or AB(80-92) results in a slight increase in Ca(2+)-affinity of recoverin and does not affect significantly a Ca(2+)-myristoyl switch mechanism of the protein. However in the presence of AB(60-72) or AB(80-92) recoverin loses its ability to interact with rhodopsin kinase and consequently to perform a function of Ca(2+)-sensitive inhibitor of rhodopsin phosphorylation in photoreceptor cells.

Mechanism of rhodopsin kinase regulation by recoverin.

Recoverin is suggested to inhibit rhodopsin kinase (GRK1) at high [Ca(2+)] in the dark state of the photoreceptor cell. Decreasing [Ca(2+)] terminates inhibition and facilitates phosphorylation of illuminated rhodopsin (Rh*). When recoverin formed a complex with GRK1, it did not interfere with the phosphorylation of a C-terminal peptide of rhodopsin (S338-A348) by GRK1.

Membrane binding of the neuronal calcium sensor recoverin - modulatory role of the charged carboxy-terminus.

Background: The Ca2+-binding protein recoverin operates as a Ca2+-sensor in vertebrate photoreceptor cells. It undergoes a so-called Ca2+-myristoyl switch when cytoplasmic Ca2+-concentrations fluctuate in the cell. Its covalently attached myristoyl-group is exposed at high Ca2+-concentrations and enables recoverin to associate with lipid bilayers and to inhibit its target rhodopsin kinase.

Tuning of a neuronal calcium sensor.

Recoverin is a Ca(2+)-regulated signal transduction modulator expressed in the vertebrate retina that has been implicated in visual adaptation. An intriguing feature of recoverin is a cluster of charged residues at its C terminus, the functional significance of which is largely unclear. To elucidate the impact of this segment on recoverin structure and function, we have investigated a mutant lacking the C-terminal 12 amino acids.

One of the Ca2+ binding sites of recoverin exclusively controls interaction with rhodopsin kinase.

Recoverin is a neuronal calcium sensor protein that controls the activity of rhodopsin kinase in a Ca(2+)-dependent manner. Mutations in the EF-hand Ca2+ binding sites are valuable tools for investigating the functional properties of recoverin. In the recoverin mutant E121Q (Rec E121Q ) the high-affinity Ca2+ binding site is disabled.