| 1995 |
RetGC-2 (GC-F/GUCY2F) was cloned from human retinal cDNA and shown to encode a membrane guanylyl cyclase expressed specifically in photoreceptor cells; recombinant RetGC-2 expressed in HEK293 cells displays guanylyl cyclase activity that is stimulated by the Ca2+-binding activator p24 (GCAP-2) and inhibited by Ca2+ with an EC50 of 50-100 nM. |
cDNA cloning, in situ hybridization, recombinant expression in HEK293 cells, in vitro guanylyl cyclase activity assay |
Proceedings of the National Academy of Sciences of the United States of America |
High |
7777544
|
| 1995 |
GC-F (GUCY2F) was isolated from a rat eye cDNA library and shown to encode a membrane guanylyl cyclase with an extracellular domain, single transmembrane domain, kinase-like domain, and catalytic cyclase domain; overexpression in COS cells conferred guanylyl cyclase activity, but known extracellular ligands for other GC receptors did not stimulate it, classifying it as an orphan receptor. |
cDNA cloning, COS cell overexpression, guanylyl cyclase activity assay, peptide ligand stimulation assay |
Proceedings of the National Academy of Sciences of the United States of America |
High |
7831337
|
| 1997 |
ROS-GC2 (the bovine ortholog of GUCY2F) was cloned from bovine retina, shown to be specifically expressed in retina, and demonstrated to be modulated in low Ca2+ by the calmodulin-like Ca2+-binding protein GCAP-2; it also contains a unique five-amino-acid signature at its C-terminus. |
cDNA cloning, retinal expression analysis, in vitro reconstitution with GCAP-2, guanylyl cyclase activity assay |
Biochemical and biophysical research communications |
High |
9175772
|
| 1998 |
The kinase homology domain (KHD) of RetGC-2 specifies the affinity and cooperativity of interaction with GCAP-2: RetGC-2 interacts cooperatively and with high affinity with GCAP-2, in contrast to RetGC-1 which interacts noncooperatively with low affinity; this was mapped using RetGC-1/RetGC-2 chimeras and a trypsin protection assay showing GCAP-2 binds constitutively to the KHD. |
Trypsin/protease protection assay, RetGC-1/RetGC-2 chimeric protein analysis, guanylyl cyclase activity assay |
Biochemistry |
High |
9698373
|
| 1998 |
CD-GCAP activates ROS-GC2 (GUCY2F ortholog), but with approximately 10-fold weaker activation than ROS-GC1; the CD-GCAP-regulated signaling switch on ROS-GC1 was mapped to amino acid segment 736-1053 through deletion mutants, hybrid constructs, and heterologous cyclase reconstruction. |
Recombinant protein expression, deletion mutagenesis, chimeric cyclase construction, in vitro guanylyl cyclase activity assay |
Biochemical and biophysical research communications |
High |
9439621
|
| 1995 |
GCAP-2 (p24) activates both RetGC-1 and RetGC-2 (GUCY2F) in a Ca2+-sensitive manner in vitro; Ca2+ inhibits activation with an EC50 near 200 nM and Hill coefficient of 1.7; recombinant GCAP-2 expressed in HEK293 cells effectively stimulates photoreceptor guanylyl cyclase. |
Protein purification, in vitro guanylyl cyclase activity assay, recombinant expression in HEK293 cells, antibody inhibition |
The Journal of biological chemistry |
High |
7559656
|
| 1999 |
GCAPs bind constitutively to an intracellular domain of RetGC-2 (GUCY2F); in the absence of Ca2+, GCAP stimulates cyclase activity, and in the presence of Ca2+, it inhibits cyclase activity; proper RetGC and GCAP functioning is necessary for photoreceptor viability. |
Biochemical binding assays, in vitro guanylyl cyclase activity assay, genetic evidence from disease mutations |
Methods (San Diego, Calif.) |
High |
10581151
|
| 1999 |
GCAP-2 functional domain mapping demonstrated that substituting specific domains of GCAP-2 with corresponding fragments from non-RetGC-regulating proteins (neurocalcin, recoverin) abolishes its ability to regulate RetGC-2 (GUCY2F) expressed in HEK293 cells; three regions are essential: residues 78-113 (determines Ca2+ activation vs. inhibition polarity), residues 29-48 (EF-1 motif, required for both activation and inhibition), and residues 171-189 (contributes to activation). |
GCAP-2 deletion mutants, chimeric protein construction, in vitro guanylyl cyclase activity assay with recombinant RetGC-1 and RetGC-2 in HEK293 cells |
The Journal of biological chemistry |
High |
10196158
|
| 1999 |
Neurocalcin, a Ca2+-binding protein structurally related to GCAPs, stimulates ROS-GC1 in a Ca2+-dependent manner (EC50 ~20 µM) but does not influence the activity of ROS-GC2 (GUCY2F ortholog), demonstrating substrate specificity between the two retinal cyclases for this regulator. |
Recombinant protein expression, in vitro guanylyl cyclase activity assay, domain mapping with deletion mutants |
Biochemistry |
Medium |
10504230
|
| 1999 |
ROS-GC2 (GUCY2F) was not detected in the pineal gland (unlike ROS-GC1), establishing that the two retinal cyclases have distinct expression patterns beyond the retina; the alpha2D/A-adrenergic receptor-linked signaling system in pinealocytes uses exclusively ROS-GC1 and not ROS-GC2. |
Immunohistochemistry, molecular cloning, immunoblotting, biochemical fractionation |
Biochemistry |
Medium |
10821676
|
| 1999 |
Disruption of GC-E (RetGC1/GUCY2D) gene in mice left GC-F (GUCY2F) expression unchanged, demonstrating that GC-F is expressed independently; however, cones progressively disappeared by 5 weeks, establishing that GC-E (not GC-F) is essential for cone photoreceptor survival. |
Gene knockout mouse, electroretinography, retinal histology, immunoblotting |
The Journal of neuroscience : the official journal of the Society for Neuroscience |
High |
10407028
|
| 2007 |
Double knockout of Gucy2e and Gucy2f (GC1/GC2) prevented transport of rod PDE6 to rod outer segments, while single Gucy2f knockout alone did not prevent transport of transducin, GRK1, or rhodopsin to rod outer segments; this established that GC-bearing membranes co-transport peripheral membrane proteins in vesicles and that GC1 and GC2 have overlapping roles in rod outer segment protein transport. |
Gucy2e/Gucy2f double knockout mice, immunofluorescence, subcellular protein localization assays |
Vision research |
High |
17949773
|
| 2009 |
GC1 (GC-E) undergoes autophosphorylation at residues Ser-530, Ser-532, Ser-533, and Ser-538 within the kinase homology domain in vivo (light- and signal transduction-independent); mutations in the putative Mg2+ binding site of the KH domain abolished phosphorylation and dramatically reduced GC activity, demonstrating that a functional KH domain is essential for cGMP production. Autophosphorylation itself does not regulate GC1 activity. |
Mass spectrometry phosphorylation mapping, site-directed mutagenesis of KH domain, in vitro guanylyl cyclase activity assay |
The Journal of biological chemistry |
High |
19901021
|
| 2015 |
ROS-GC2 (GUCY2F ortholog) is activated by bicarbonate in a Ca2+- and GCAP-independent manner (ED50 ~39 mM), with bicarbonate stimulation being more powerful in the presence of GCAP1 or GCAP2 at low Ca2+; this reveals a novel regulatory input to the ROS-GC system beyond Ca2+/GCAP signaling. |
Recombinant protein guanylyl cyclase activity assay, photoreceptor electrophysiology (circulating current and flash response measurements) |
The Journal of biological chemistry |
High |
25767116
|
| 2018 |
RD3 (retinal degeneration protein 3) inhibits both GC-E and GC-F (GUCY2F) and is involved in their transport from inner to outer segments; additionally, RD3 directly interacts with guanylate kinase (demonstrated by back-scattering interferometry) and co-localizes with guanylate kinase in photoreceptor inner segments, revealing a role in the nucleotide cycle. |
Back-scattering interferometry (direct interaction assay), immunohistochemistry, guanylyl cyclase activity assay, mouse retina sections |
Frontiers in molecular neuroscience |
Medium |
29515371
|
| 2012 |
Gucy2f knockdown in zebrafish using morpholino oligonucleotides (blocking translation or splicing) resulted in significantly reduced visual function (optomotor response) and histological changes including loss and shortening of cone and rod outer segments, establishing GUCY2F as required for photoreceptor outer segment integrity and visual function in vivo. |
Zebrafish morpholino knockdown, optomotor assay, retinal histology |
European journal of human genetics : EJHG |
Medium |
22378290
|
| 2009 |
Overexpression of gucy2F throughout the zebrafish nervous system via Gal4-UAS system caused multiple defects including loss of forebrain neurons, demonstrating that proper control of cGMP production by GUCY2F is important for neuronal survival. |
Zebrafish gain-of-function screen using MMLV insertion with Gal4-UAS overexpression system, histological analysis of forebrain neurons |
Molecular genetics and genomics : MGG |
Medium |
19221799
|
| 1996 |
The GC-F gene (Gucy2f) was mapped to the X chromosome in mice (chromosome X) and the human homolog was localized to Xq22 by fluorescence in situ hybridization; genomic organization analysis showed conservation of exon-intron boundaries with other guanylyl cyclase genes. |
Mouse interspecific backcross analysis, fluorescence in situ hybridization (FISH), genomic library screening |
Genomics |
Medium |
8838319
|
| 2023 |
Evolutionary analysis across vertebrates showed that GC-F (GUCY2F) is absent in several clades (reptiles, birds, marsupials); in species lacking GC-F, visual function is compensated by an increased number of GCAPs, while in nocturnal/visually impaired species, GC-F loss is paralleled by GCAP inactivation, indicating GC-E and GC-F together regulate phototransduction cGMP synthesis with partially compensatory roles. |
Comparative genomics, phylogenetic analysis across vertebrate species |
Frontiers in molecular neuroscience |
Low |
37007786
|
| 2023 |
In an AAV-CRISPR 'ablate and replace' study, Gucy2f-/- mice served as a model where loss of GC-F alone allowed testing of GUCY2D (GC-E/RetGC1) rescue vectors; Gucy2e+/-:Gucy2f-/- mice were used to optimize vector doses for CORD6 therapy, confirming that GC-F contributes to but is not solely required for photoreceptor function when GC-E is partially present. |
Gucy2f knockout mice, AAV-CRISPR-Cas9 delivery, electroretinography, retinal structure analysis |
Molecular therapy. Methods & clinical development |
Medium |
37361352
|