Retina International's Scientific Newsletter - Guanylate Cyclase Activating Protein

	Retina International's Scientific Newsletter
Editor's Notes from literature on Guanylate Cyclase Activating Protein (GCAP)

Recent update from: 19.12.1999

GCAP

Notes

GCAP is myristoylated

Species:

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (6)

GCAP

Notes

IR of GCAP1 in CIS, COS, and axons as well as RIS with lower IR, very low in ROS
In mouse GCAP1-IR only in COS, in rat only in CIS
IR of GCAP2 in man and monkey strong in COS, in bovine weak, in rodents faint
In primates GCAP2-IR was strong in RIS and faint in ROS
In cat, mouse, rat strong in RIS and ROS
GCAP2-IR evident in amacrines, bipolars, and ganglion cells
GCAP1 and 2 in synaptic termini in rods and cones

Species:

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (4)

GCAP

Notes

GCAP1 in COS and ROS activate RetGC (outer segment GC) and endogenous GC at low Ca²⁺
GCAP2 50% identical to GCAP1
GCAP2 activates RetGC in inner retina/photoreceptor inner segment
RetGC is outer segment GC

Species:

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (12)

GCAP

Notes

IR of GCAP1 in CIS, COS, and axons as well as rods with lower IR
In mouse GCAP1-IR only in COS, in rat only in CIS
IR of GCAP2 in man strong COS, in bovine weak, in rodents faint
In cat, rat, mouse in rods

Species:

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (5)

GCAP1

Notes

Only 3 of the 4 EF hands bind Ca²⁺
EF1 alteration does not affect Ca²⁺ binding
In Ca²⁺-bound state only C- and N-terminus are removed by trypsin
Without Ca²⁺ GCAP1 is degraded by trypsin into small fragments
Only EF3/EF4 contribute to Ca²⁺ inactivation of GC => major conformational changes during Ca²⁺ binding

Species: Bos taurus

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (21)

GCAP1

Notes

Cyclase activity when Ca²⁺ is lowered to 45 nM
Monomer

Species: Bos taurus

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE: 20 kDa
	calculated: 23 kDa

Reference: (11)

GCAP

Notes

RNA only in inner segments
35% homology to recoverin
3 Ca²⁺ binding sites (EF-hand)
possible phenotype with progressive CSNB
soluble acidic protein, lacking hydrophobic domains
N-terminal acylation at Gly2
possible N-terminal palmitoylation at Cys18
residues 1-180 conserved in bovine, mouse, and man

Species: Homo sapiens

See also:

Chromosomal localisation	in man: 6p21.1
	in mouse:

Gene Data

Exons: 4

Primers:

Protein Data

Aminoacids: 201 AA

Molecularweight	on SDS-PAGE: 21-23 kDa
	calculated:

Reference: (20)

GCAP1

Notes

Species: Homo sapiens

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids: 205 AA

Molecularweight	on SDS-PAGE: 20 kDa
	calculated: 20594

Reference: (2)

GCAP

Notes

Both GCAP stimulated ROS-GC at low Ca²⁺
Two different binding sites suggested for GACP1 and 2
Deletion of extracellular domain does not block activity of ROS-GC

Species: Homo sapiens

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (10)

GACP

Notes

GCAPs are arranged in a tail-to-tail array
Diverged from mouse more than 300 mio years ago

Species: Homo sapiens

See also:

Chromosomal localisation	in man: 6p
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (14)

GCAP2

Notes

2 GCAP: GCAP1 = GCAP
GCAP2 = P24 regulates RetGC2 not RetGC1
Ca²⁺ type protein in retina and outer segment preparations
Strong rod signals
Maybe masked epitopes in cones
Ca²⁺ elutes GCAP2 from membranes

Species: Homo sapiens

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (9)

GCAP2

Notes

Membrane association is Ca²⁺-dependent not myristoylation- or acylation-dependent
Myristoylation does not effect RetGC activation

Species: Homo sapiens

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (8)

GACP

Notes

GCAPs are arranged in a tail-to-tail array

Species: Mus musculus

See also:

Chromosomal localisation	in man:
	in mouse: 17 D

Gene Data

gDNA: <18 kb

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (14)

GACP1

Notes

Species: Mus musculus

See also:

Chromosomal localisation	in man:
	in mouse: 17 D

Gene Data

mRNA: 0.8 kb

Exons: 4

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (14)

GACP2

Notes

Species: Mus musculus

See also:

Chromosomal localisation	in man:
	in mouse: 17 D

Gene Data

mRNA: 2.0 kb

Exons: 4

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (14)

GCAP

Notes

Species: Mus musculus

See also:

Chromosomal localisation	in man:
	in mouse: 17

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (3)

GCAP

Notes

Head to tail tandem as in mammals

Species: Rana pipiens

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids: 205 AA

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (22)

GCAP

Notes

Frog GCAP regulates human RetGC1
ROSGC = RetGC1
N-terminal myristoylation consensus sequence

Species: Rana?

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids: 205 AA

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (19)

GCAP, GCAP1

Notes

Y99C mutant alters Ca²⁺ sensitivity considerably resulting in persistent stimulation of GC1
Mutations might impair the production of cGMP thus producing permanent closure of CNGA1
Mutations do not influence retinal development and unusual fundus changes are absent
WT and mutant
- are acylated at a similar amount
- are similar effective for the stimulation of GC1
Mutation causes reduced but persistent stimulation of GC1

Species:

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (18)

GCAP, retinal

Notes

Binds 2 - 3 Ca²⁺
Activates Guanylate cyclase
Pi = 4,1
N-acetylated
Affects GC at low Ca²⁺

Species: Bos taurus

See also: GUC

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids: 180 AA

Molecularweight	on SDS-PAGE:
	calculated: 20594

Reference: (2)

GCAP2, rod

Notes

fatty acylated
EF1 is not a real EF-hand

Species: Homo sapiens

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (7)

GUCA1A, rod/cone

Notes

Mutations underlying ad cone rod dystrophy

Species: Homo sapiens

See also:

Chromosomal localisation	in man: 6p21.1
	in mouse:

Gene Data

Exons: 4

Primers: complete

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (15)

GCAP1, type 1

Notes

Homolgy to vertebrate GCP 78-80%

Species: G_allus gallus

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids: 199 AA

Molecularweight	on SDS-PAGE:
	calculated: 22810

Reference: (16)

GCAP1, type 1

Notes

Transcripts are reduced to 70% in rd/rd chicken
GCAP1 levels are reduced by 90%
Met1 is posttranslationally cleaved
Only 43% similarity to GCAP2, to bovine GCAP1 79%
Contains 3 EF-hands

Species: G_allus gallus

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids: 199 AA

Molecularweight	on SDS-PAGE:
	calculated: 22822

Reference: (17)

GCAP1, type 1

Notes

No mutations found

Species: Homo sapiens

See also:

Chromosomal localisation	in man: 6p21.1
	in mouse:

Gene Data

Exons: 4

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (1)

Gcap, type 1

Notes

RNA only in inner segments
35% homology to recoverin
3 Ca²⁺ binding sites (EF-hand)
Possible phenotype with progressive CSNB
Soluble acidic protein, lacking hydrophobic domains
N-terminal acylation at Gly2
Possible N-terminal palmitoylation at Cys18
Residues 1-180 conserved in bovine, mouse, and man

Species: Mus musculus

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons: 4

Primers:

Protein Data

Aminoacids: 202 AA

Molecularweight	on SDS-PAGE: 21-23 kDa
	calculated:

Reference: (20)

Gcap1, type 1

Notes

Expressed in retinal photoreceptors
Tail to tail array with Gcap 2

Species: Mus musculus

See also:

Chromosomal localisation	in man:
	in mouse: 17D

Gene Data

mRNA: 0.8 kb

Exons:

Primers:

Protein Data

Aminoacids: 202 AA

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (13)

GCAP2, type 2

Notes

Transcripts are reduced to 70% in rd/rd chicken
GCAP2 levels are normal while
Met1 is posttranslationally cleaved
Similarity to bovine GCAP2 77%
Contains 3 EF-hands

Species: G_allus gallus

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids: 198 AA

Molecularweight	on SDS-PAGE:
	calculated: 23128

Reference: (17)

GUCA, type 2

Notes

2 GCAP: GCAP1 and GCAP2 = p24
GCAP1 in COS
3 EF-hands

Species: Homo sapiens

See also:

Chromosomal localisation	in man: 6
	in mouse: 17

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (3)

Gcap2, type 2

Notes

Expressed in retinal photoreceptors but not in cone
Expressed in amacrine and ganglion cells
Tail to tail array with Gcap 1

Species: Mus musculus

See also:

Chromosomal localisation	in man:
	in mouse: 17D

Gene Data

mRNA: 2 kb

Exons:

Primers:

Protein Data

Aminoacids: 201 AA

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (13)

References:

1. Amberson, D.A., Baehr, W., Subbaraya, I., Berson, E.L., and Dryja, T.P. Screen for mutations in the gene encoding guanylate cyclase activating protein in patients with retinitis pigmentosa. 1995; Invest.Ophthalmol.Vis.Sci. Suppl. S826
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2. Baehr, W., Gorczyca, W., Subbaraya, I., Ohguro, H., Johnson, R.S., Walsh, K.A., Gray Keller, M.P., Detwiler, P.B., Crabb, J., and Palczewski, K. Isolation, cloning, and functional characterization of bovine retina guanylate cyclase activating protein (GCAP). 1994; Invest.Ophthalmol.Vis.Sci. 35 (Suppl.): 1486
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3. Baehr, W., Subbaraya, I., Dang, Y.L., Ruiz, C., Helekar, B., Banerjee, P., Knowles, J.A., Gilliam, T.C., Vandenburgh, K., and Stone, E.M. Human and mouse GCAP genes and involvement of GCAP gene mutations in human disease. 1995; Invest.Ophthalmol.Vis.Sci. 36: S826
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4. Cuenca, N., Lopez, S., Howes, K., and Kolb, H. The localization of guanylyl cyclase-activating proteins in the mammalian retina. 1998; Invest.Ophthalmol.Vis.Sci. 39: 1243 - 1250.
Goto Top Link to PudMed

5. Cuenca, N., Wall, S., Lopez, S., Howes, K., Baehr, W., and Kolb, H. Immunocytochemical Localization Of Guanylate Cyclase activating Protein (GCAP1) To The Neurons Of The Mammalian Retina. 1997; Invest.Ophthalmol.Vis.Sci. 38: S21 - A1000.
Goto Top

6. Demar, J.C., Wensel, T.G., and Anderson, R.E. Metabolic pathways for the generation of the unsaturated C-14 fatty acids found on the N-terminus of transducin. 1995; Invest.Ophthalmol.Vis.Sci. 36: S223
Goto Top

7. Dizhoor, A.M., Olshevskaya, E.V., Ankoudinova, I., and Hurley, J.B. Role Of The N-Terminus And Calcium Binding Domains Of GCAP-2 In Regulating Photreceptor Guanylyl Cyclase. 1996; Invest.Ophthalmol.Vis.Sci. 37: S215
Goto Top

8. Dizhoor, A.M., Olshevskaya, E.V., Hughes, R., and Hurley, J.B. The N Terminal Fatty Acyl Group Of GCAP2 Is Not Essential For Guanylyl Cyclase Regulation Or For Membrane Binding. 1997; Invest.Ophthalmol.Vis.Sci. 38: S477 - A1000.
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9. Dizhoor, A.M., Olshevskaya, E.V., and Hurley, J.B. A novel retinal Ca2+-binding protein, p24, stimualtes photoreceptor guanylyl cyclase and associates with the outer segment membranes in a calcium-sensitive manner. 1995; Invest.Ophthalmol.Vis.Sci. 36: S223
Goto Top

10. Duda, T., Krishnan, A., Goraczniak, R.M., and Sharma, R.K. Mechanisms Of GCAP1 And GCAP2 Activation Of ROS-GC. 1997; Invest.Ophthalmol.Vis.Sci. 38: S21 - A1000.
Goto Top

11. Gorczyca, W.A., Gray Keller, M.P., Detwiler, P.B., and Palczewski, K. Purification and physiological evaluation of a guanylate cyclase activating protein from retinal rods Purification and physiological evaluation of a guanylate cyclase activating protein from retinal rods. 1994; Proc.Natl.Acad.Sci.U.S.A. 91: 4014 - 4018.
Goto Top Link to PudMed

12. Gorczyca, W.A., Polans, A.S., Surgucheva, I.G., Subbaraya, I., Baehr, W., and Palczewski, K. Guanylyl cyclase activating protein - A calcium-sensitive regulator of phototransduction. 1995; J.Biol.Chem. 270: 22029 - 22036.
Goto Top

13. Howes, K., Bronson, J.D., Dang, Y.L., Li, N., Zhang, K., Ruiz, C., Helekar, B., Lee, M., Subbaraya, I., Kolb, H., Chen, J., and Baehr, W. Gene array and expression of mouse retina guanylate cyclase activating proteins 1 and 2. 1998; Invest.Ophthalmol.Vis.Sci. 39: 867 - 875.
Goto Top Link to PudMed

14. Howes, K., Bronson, J.D., Li, N., Zhang, K., Lee, M., Subbaraya, I., Frederick, J., Surguchov, A., Kolb, H., Palczewski, K., and Baehr, W. Molecular Characterization Of The Mouse GCAP1 GCAP2 Gene Array And Expression Of Gcaps In The Mouse Retina. 1997; Invest.Ophthalmol.Vis.Sci. 38: S21 - A1000.
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15. Payne, A.M., Dowes, S.M., Bessant, D.A.R., Taylor, R., Holder, G.E., Warren, M.J., Bird, A.C., Bhattacharya, S.S., Downes, S.M., and Bessant, D.A. A mutation in guanylate cyclase activator 1A (GUCA1A) in an autosomal dominant cone dystrophy pedigree mapping to a new locus on chromosome 6p21.1. 1998; Hum.Mol.Genet. 7: 273 - 277.
Goto Top Link to PudMed

16. Semple Rowland, S.L., Helekar, B., Ruiz, C., Zhao, X., Palczewski, K., and Baehr, W. Normal and rd/rd mutant chicken retina guanylate cyclase-activating protein (GCAP). 1995; Invest.Ophthalmol.Vis.Sci. 36: S222
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17. Semple-Rowland, S., Buczylko, J., Palczewski, K., and Baehr, W. Abnormal Expression Of GCAP1 And Photoreceptor Guanylate Cyclase In rd Chicken Retina. 1996; Invest.Ophthalmol.Vis.Sci. S810
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18. Sokal, I., Li, N., Surgucheva, I., Warren, M.J., Payne, A.M., Bhattacharya, S.S., Baehr, W., and Palczewski, K. GCAP1 (Y99C) mutant is constitutively active in autosomal dominant cone dystrophy. 1998; Mol.Cell. 2: 129 - 133.
Goto Top Link to PudMed

19. Subbaraya, I., Ruiz, C., and Baehr, W. Cloning and expression of frog (Rana pipiens) guanylyl cyclase activating protein (GCAP). 1995; Invest.Ophthalmol.Vis.Sci. 36: S223
Goto Top

20. Subbaraya, I., Ruiz, C.C., Helekar, B.S., Zhao, X., Gorczyca, W.A., Pettenati, M.J., Rao, P.N., Palczewski, K., and Baehr, W. Molecular characterization of human and mouse photoreceptor guanylate cyclase-activating protein (GCAP) and chromosomal localization of the human gene. 1994; J.Biol.Chem. 269: 31080 - 31089.
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21. Surgucheva, I., Rudnicka Nawrot, M., Hulmes, J.D., Haeseleer, F., Sokal, I., Crabb, J.W., Baehr, W., and Palczewski, K. Bovine Photoreceptor GCAP1: Changes of Biological Activity And Structure As A Function of Ca++. 1998; Invest.Ophthalmol.Vis.Sci. 39: S951
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22. Zhang, K., Ferraro, A., Kniazeva, M., Han, M., and Donoso, L.A. Clinical Characteristics Of Autosomal Dominant Stargardt's Macular Dystrophy With Genetic Linkage To Chromosome 13q34. 1997; Invest.Ophthalmol.Vis.Sci. 38: S959 - A1000.
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Contact the editor: irpamp@irpa.org	This site is maintained and edited by Dr. rer. medic. Markus Preising, Dipl.Biol. Molecular Genetics Laboratory Department of Paediatric Ophthalmology, Strabismology and Ophthalmogenetics University of Regensburg Head: Prof. Dr. med. Birgit Lorenz