Ca(2+) sensor S100beta-modulated sites of membrane guanylate cyclase in the photoreceptor-bipolar synapse.
This study documents the identity of a calcium- regulated membrane guanylate cyclase transduction system in the photoreceptor-bipolar synaptic region. The guanylate cyclase is the previously characterized ROS-GC1 from the rod outer segments and its modulator is S100beta. S100beta senses increments in free Ca(2+) and stimulates the cyclase. Specificity of photoreceptor guanylate cyclase activation by S100beta is validated by the identification of two S100beta-regulatory sites. A combination of peptide competition, surface plasmon resonance binding and deletion mutation studies has been used to show that these sites are specific for S100beta and not for another regulator of ROS-GC1, guanylate cyclase-activating protein 1. One site comprises amino acids (aa) Gly962-Asn981, the other, aa Ile1030-Gln1041. The former represents the binding site. The latter binds S100beta only marginally, yet it is critical for control of maximal cyclase activity. The findings provide evidence for a new cyclic GMP transduction system in synaptic layers and thereby extend existing concepts of how a membrane-bound guanylate cyclase is regulated by small Ca(2+)-sensor proteins.
Duda
Teresa
T
The Unit of Regulatory and Molecular Biology, Departments of Cell Biology and Ophthalmology, NJMS & SOM/UMDNJ, Stratford, NJ 08084, USA.
Koch
Karl-Wilhelm
KW
Venkataraman
Venkateswar
V
Lange
Christian
C
Beyermann
Michael
M
Sharma
Rameshwar K
RK
D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D015153:Blotting, Western; D019556:COS Cells; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D002417:Cattle; D004305:Dose-Response Relationship, Drug; D017353:Gene Deletion; D006162:Guanylate Cyclase / Q000737:chemistry* / Q000378:metabolism; D007150:Immunohistochemistry; D009414:Nerve Growth Factors / Q000737:chemistry* / Q000378:metabolism*; D009842:Oligopeptides / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D010786:Photoreceptor Cells / Q000201:enzymology*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012160:Retina / Q000201:enzymology; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins; D020349:Surface Plasmon Resonance; D013569:Synapses / Q000378:metabolism*; D013997:Time Factors
D002135:Calcium-Binding Proteins; D009414:Nerve Growth Factors; D009842:Oligopeptides; D010455:Peptides; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins; D006162:Guanylate Cyclase; D002118:Calcium