Guanylate-cyclase-inhibitory protein is a frog retinal Ca2+-binding protein related to mammalian guanylate-cyclase-activating proteins.
Two guanylate-cyclase-activating proteins (GCAP) encoded by a tail-to-tail gene array have been characterized in the mammalian retina. Using frog retina as a model, we obtained evidence for the presence of a photoreceptor Ca2+-binding protein closely related to GCAP. This protein (206 amino acids) does not stimulate guanylate cyclase (GC) in low [Ca2+], but inhibits GC in high [Ca2+], and is therefore termed guanylate-cyclase-inhibitory protein (GCIP). Sequence analysis indicates that GCIP and GCAP1 and GCAP2 have diverged substantially, but conserved domains present in all vertebrate GCAP are present in GCIP. Moreover, partial characterization of the GCIP gene showed that the positions of two introns in the GCIP gene are identical to positions of corresponding introns of the mammalian GCAP gene array. As to the major differences between GCIP and GCAP, the fourth EF hand Ca2+-binding motif of GCIP is disabled for Ca2+ binding, and GCIP does not stimulate GC. Monoclonal and polyclonal antibodies raised against recombinant GCIP identified high levels of GCIP in the inner segments, somata and synaptic terminals of frog cone photoreceptors. The results suggest that GCIP is a Ca2+-binding protein of the GCAP/recoverin subfamily. Its localization in frog cones closely resembles that of GC in mammalian cones. GCIP inhibits GC at high free [Ca2+], competing with GCAP1 and GCAP2 for GC regulatory sites.
Li
N
N
Moran Eye Center, University of Utah Health Science Center, Salt Lake City 84132, USA.
Fariss
R N
RN
Zhang
K
K
Otto-Bruc
A
A
Haeseleer
F
F
Bronson
D
D
Qin
N
N
Yamazaki
A
A
Subbaraya
I
I
Milam
A H
AH
Palczewski
K
K
Baehr
W
W
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D002417:Cattle; D003001:Cloning, Molecular; D017931:DNA Primers; D015723:Gene Library; D051604:Guanylate Cyclase-Activating Proteins; D008322:Mammals; D008969:Molecular Sequence Data; D010786:Photoreceptor Cells / Q000378:metabolism; D017433:Protein Structure, Secondary; D011894:Rana pipiens; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012160:Retina / Q000378:metabolism*; D012374:Rod Cell Outer Segment / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D002135:Calcium-Binding Proteins; D017931:DNA Primers; D051604:Guanylate Cyclase-Activating Proteins; D011994:Recombinant Proteins