Retina International's Scientific Newsletter

	Retina International's Scientific Newsletter
Editor's Notes from literature on Recoverin

Recent update from: 19.12.1999

RCV

Notes

Interacts with IRBP, RHOK and tubulin in the presence of Ca²⁺
Introduces Ca²⁺ sensitivity to RHOK
mristoylation introduces cooperativity to Ca²⁺ sensitivity

Species: Bos taurus

See also: RHOK

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (4)

Rcv

Notes

EF-hand Ca²⁺-binding motifs
Homology to visinin
Each EF-hand comprises of 29 AA

Species: Bos taurus

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids: 202 AA

Molecularweight	on SDS-PAGE: 26 kDa
	calculated:

Reference: (12)

RCV

Notes

mristoalyted by N-mristoyltransferase (EC 2.3.1.97)
N-terminal Met is posttranslationally removed
Cristallographic data

Species: Bos taurus

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids: 202 AA

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (21)

RCV

Notes

pI = 5.25
Ca²⁺-dependent mobility shift in a native gel
Monomeric
Protein of EF-hand family
Phosphorylated at low [Ca²⁺]
Ratio RCV:RHO = 1:160

Species: Bos taurus

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (13)

RCV

Notes

Ca²⁺ binding protein
Heterogeneously acylated at N-terminal Gly
Most abundant acyl group: mristoleate (14:1)
Also: 14:0, 14:2, 12:0
N-terminus matches consensus sequence for N-mristoyl-transferase (GNSKS)

Species: Bos taurus

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated: 23,412.2

Reference: (7)

RCV

Notes

Does not activate Guanlate Cyclase
mristoylated
Dissociates from membranes at low [Ca²⁺]
Raised RCV concentration slows recovery from photoexcitation

Species: Bos taurus

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE: 23 kDa
	calculated:

Reference: (11)

RCV

Notes

Promotes recovery of the dark state
3 potential EF-hands
Not detected in brain, heart, liver, lung, kidney
IR in the entire photoreceptor layer
Modified N-terminus
Identity to visinin 59.9%
Binds Ca²⁺ at low concentration
GC activation with variable results =? unstable preparation or additional factors

Species: Bos taurus

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids: 202AA

Molecularweight	on SDS-PAGE: 26 kDa
	calculated: 23.3 kDa

Reference: (8)

RCV

Notes

Binds RHOK

Species: Bos taurus

See also: RHOK

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (3)

RCV

Notes

Ca²⁺ changes a-helix amount from 38% to 50%
Na⁺ affects attachment to membrane
Fatty acid modification

Species: Bos taurus

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (20)

RCV

Notes

mristoylation is necessary for membrane attachment
mristoylation is not necessary for Ca²⁺ dependent RHOK inhibition
mristoylation induces a cooperative dependence of RHOK inhibition

Species: Bos taurus

See also: RHOK

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (2)

RCV

Notes

Microsatellite RCV1: (CA)7TG(CA)5(CG)4CA9 in 3' UTR
Alleles RCV-14CA: A1=198 bp (11%), A2=196 (10%), A3=194 bp (18%), A4=192 bp (1%), A5=190 bp (60%); PIC: 0.59
Alleles RCV-3PRCA: B1=208 bp (18%), B2=206 bp (3%), B3=200 bp (79%); PIC: 0.34
Microsatellite D17S945: Genebank: Z24012
Microsatellite RCV-14CA: Genebank: Z24012
Between MH2 and D17S1

Species: Homo sapiens

See also:

Chromosomal localisation	in man: 17p13
	in mouse:

Gene Data

Exons:

Primers: Repeat

Protein Data

Aminoacids: 200 AA

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (22)

RCV

Notes

Species: Homo sapiens

See also:

Chromosomal localisation	in man: 17p13.1
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (14)

RCV

Notes

Silent mutations:
Ser 214 Ser
Pro 190 Pro
Non pathogenic Ala 200 Thr

Species: Homo sapiens

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers: Man

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (18)

RCV

Notes

IR in photoreceptors and in a subpopulation of bipolars
Very robust labeling in a small population of cells in the GCL
Strong sequence homology to neurocalcin
IR-positive bipolars are cone bipolars in human, monkey, and rat
IR in photoreceptors was very strong in inner segments
Expression in human retina without specific localization
Rods and cones show hybridization with riboprobe

Species: Homo sapiens

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (23)

RCV

Notes

58% homology to visinin, 87% homology to bovine recoverin
9-10 kb genomic DNA
3 EF-hands
Mediates recovery of the dark

Species: Homo sapiens

See also:

Chromosomal localisation	in man: 17
	in mouse:

Gene Data

Exons: 3

Primers:

Protein Data

Aminoacids: 200 AA

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (17)

RCV

Notes

no involvement of RCV in spanish ARRP patients
Microsattelite: (CA)7TG(CA)4(CA)9 in 3' UTR, 3 alleles: 150 bp, 148 bp, 142 bp, PIC: 0.46
Linked marker: D17S786, flanking: D17S796, D17S804
9 - 10 kb single-copy gene

Species: Homo sapiens

See also:

Chromosomal localisation	in man: 17p13
	in mouse:

Gene Data

Exons: 3

Primers: Complete exons, Microsattelite

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (1)

RCV

Notes

Species: Homo sapiens

See also:

Chromosomal localisation	in man: 17p13.1
	in mouse:

Gene Data

cDNA:

Exons: 3

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated: 23 kDa

Reference: (10)

RCV

Notes

Silent mutations:
- Ser 24 (TCG - TCC) frequency 0.5%
- Val 122 (GTG - GTA)
- IVS2-11c-g (heterozygous in USHII patient
- No cosegregation Ala 200 Thr Frequency 0.2%
596 individuals examined
Exon-intron boundaries

Species: Homo sapiens

See also:

Chromosomal localisation	in man: 17p13.1
	in mouse:

Gene Data

Exons:

Primers: Complete

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (19)

RCV

Notes

88% AA identity to bovine RCV
Inhibited at high [Ca²⁺]
Homologous to chicken visinin
IR in photoreceptors including outer segments
IR in INL in bipolars
Highly conserved EF-hands

Species: Homo sapiens

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

cDNA: 1100 bp

Exons:

Primers:

Protein Data

Aminoacids: 200 AA

Molecularweight	on SDS-PAGE:
	calculated: 23.1 kDa

Reference: (24)

RCV

Notes

FISH mapping
(CA)7TG(CA)5(CG)4(CA)9
Flanking markers: D17S1 - MH2

Species: Homo sapiens

See also:

Chromosomal localisation	in man: 17p13 distal
	in mouse:

Gene Data

Exons:

Primers: Repeat

Protein Data

Aminoacids: 200 AA

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (22)

Rcv

Notes

Present only in rods (rodless adult rd mice do not stain)
pI=4.83
3 potential PRKC phosphorylation sites
2 EF-hand Ca²⁺-binding domains
Phosphorylation by an endogenous kinase unequal to RHOK or PRKC
Potential glycosylation site at Gly 113
Possible regulation by glycosylation

Species: Mus musculus

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

mRNA: 1.0 kb

Exons:

Primers:

Protein Data

Aminoacids: 202 AA

Molecularweight	on SDS-PAGE: 23 kDa
	calculated: 23406 Da

Reference: (16)

RCV

Notes

RCV knock out mouse
Inhibition of RHO phosphorylation followed by prolonged PDE activation
No degeneration, no morphological changes
ERGs appear normal

Species: Mus musculus

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (5)

RCV

Notes

Species: Mus musculus

See also:

Chromosomal localisation	in man:
	in mouse: 11

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (6)

Rcvrn

Notes

Species: Mus musculus

See also:

Chromosomal localisation	in man:
	in mouse: 11 Sparc-Zfp-3

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (15)

RCV

Notes

RCV knock-out mouse
Impaired adaptation
Reduced Ca²⁺ buffer capacity

Species: Mus musculus

See also:

Chromosomal localisation	in man:
	in mouse:

Gene Data

Exons:

Primers:

Protein Data

Aminoacids:

Molecularweight	on SDS-PAGE:
	calculated:

Reference: (9)

References:

1. Bayes, M., Valverde, D., Balcells, S., Grinberg, D., Vilageliu, L., Benitez, J., Ayuso, C., Beneyto, M., Baiget, M., and Gonzalez Duarte, R. Evidence against involvement of recoverin in autosomal recessive retinitis pigmentosa in 42 Spanish families. 1995; Hum.Genet. 96: 89 - 94.
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2. Calvert, P.D., Klenchin, V.A., and Bownds, M.D. Rhodopsin Kinase Autophosphorylation Regulates It's Sensivity To Calcium-Recoverin. 1995; Invest.Ophthalmol.Vis.Sci. 36: S269
Goto Top

3. Chen, C.K. and Hurley, J.B. Calcium dependent recoverin/rhodopsin kinase interaction. 1994; Invest.Ophthalmol.Vis.Sci. 35: 1485
Goto Top

4. Chen, C.K., Inglese, J., Lefkowitz, R.J., and Hurley, J.B. Ca(2+)-dependent interaction of recoverin with rhodopsin kinase. 1995; J.Biol.Chem. 270: 18060 - 18066.
Goto Top

5. Chen, Z., Prasad, S., and Cynader, M.S. Establishment of cDNA libraries of twelve different human ocular tissues. 1995; Invest.Ophthalmol.Vis.Sci. Suppl. S123
Goto Top

6. Danciger, M., Kozak, C.A., Applebury, M.L., Polans, A., and Farber, D.B. Chromosomal mapping in the mouse of the genes for two proteins expressed in photoreceptors: cone cGMP-PDE-alpha and recoverin. 1993; Invest.Ophthalmol.Vis.Sci. 34 (Suppl.): 1461
Goto Top

7. Dizhoor, A.M., Ericsson, L.H., Johnson, R.S., Kumar, S., Olshevskaya, E., Zozulya, S., Neubert, T.A., Stryer, L., Hurley, J.B., and Walsh, K.A. The NH2 terminus of retinal recoverin is acylated by a small family of fatty acids. 1992; J.Biol.Chem. 267: 16033 - 16036.
Goto Top Link to PudMed

8. Dizhoor, A.M., Ray, S., Kumar, S., Niemi, G., Spencer, M., Brolley, D., Walsh, K.A., Philipov, P.P., Hurley, J.B., and Stryer, L. Recoverin: a calcium sensitive activator of retinal rod guanylate cyclase. 1991; Science. 251: 915 - 918.
Goto Top

9. Dodd, R.L., Makino, C.L., Chen, J., Simon, M.I., and Baylor, D.A. Visual transduction in transgenic mouse rods lacking recoverin. 1995; Invest.Ophthalmol.Vis.Sci. 36: S641
Goto Top

10. Freund, C.L. and Valle, D. Cloning and initial characterization of the cDNA and gene for human recoverin. 1992; Am.J.Hum.Genet. 51: Abst.13.
Goto Top

11. Hurley, J.B., Dizhoor, A.M., Ray, S., and Stryer, L. Recoverin's role: conclusion withdrawn [letter; comment]. 1993; Science. 260: 740
Goto Top

12. Kutuzov, M.A., Shmukler, B.E., Suslov, O.N., Dergachev, A.E., Zargarov, A.A., and Abdulaev, N.G. P26-calcium binding protein from bovine retinal photoreceptor cells. 1991; FEBS Lett. 293: 21 - 24.
Goto Top Link to PudMed

13. Lambrecht, H.G. and Koch, K.W. Recoverin, a novel calcium-binding protein from vertebrate photoreceptors. 1992; Biochim.Biophys.Acta. 1160: 63 - 66.
Goto Top

14. McGinnis, J.F., Austin, B., Klisak, I., Heinzmann, C., Kojis, T., Sparkes, R.S., Bateman, J.B., and Lerious, V. Chromosomal assignment of the human gene for the cancer-associated retinopathy protein (recoverin) to chromosome 17p13.1. 1995; J.Neurosci.Res. 40: 165 - 168.
Goto Top Link to PudMed

15. McGinnis, J.F., Lerious, V., Pazik, J., and Elliott, R.W. Chromosomal assignment of the recoverin gene and cancer-associated retinopathy. 1993; Mammalian.Genome. 4: 43 - 45.
Goto Top

16. McGinnis, J.F., Stepanik, P.L., Baehr, W., Subbaraya, I., and Lerious, V. Cloning and sequencing of the 23 kDa mouse photoreceptor cell-specific protein. 1992; FEBS Lett. 302: 172 - 176.
Goto Top Link to PudMed

17. Murakami, A., Yajima, T., and Inana, G. Isolation of human retinal genes: recoverin cDNA and gene. 1992; Biochem.Biophys.Res.Commun. 187: 234 - 244.
Goto Top Link to PudMed

18. Parminder, A.H., Murakami, A., Inana, G., Berson, E.L., and Dryja, T.P. Screen for mutations in the human recoverin gene in patients with retinitis pigmentosa and allied diseases. 1995; Invest.Ophthalmol.Vis.Sci. 36: S893
Goto Top

19. Parminder, A.H., Murakami, A., Inana, G., Berson, E.L., and Dryja, T.P. Evaluation of the human gene encoding recoverin in patients with retinitis pigmentosa or an allied disease. 1997; Invest.Ophthalmol.Vis.Sci. 38: 704 - 709.
Goto Top Link to PudMed

20. Polans, A.S., Witkowska, D., Haley, T.L., and Crabb, J.W. Recoverin and CAPL: The role of calcium-binding proteins in transduction and ocular disease. 1994; Invest.Ophthalmol.Vis.Sci. 35: 1485
Goto Top

21. Ray, S., Zozulya, S., Niemi, G.A., Flaherty, K.M., Brolley, D., Dizhoor, A.M., McKay, D.B., Hurley, J., and Stryer, L. Cloning, expression, and crystallization of recoverin, a calcium sensor in vision. 1992; Proc.Natl.Acad.Sci.U.S.A. 89: 5705 - 5709.
Goto Top

22. Wiechmann, A.F. Three microsatellite polymorphisms at the recoverin locus on chromosome 17. 1994; Hum.Mol.Genet. 3: 1028
Goto Top

23. Wiechmann, A.F. and Hammarback, J.A. Expression of recoverin mRNA in the human retina: localization by in situ hybridization. 1993; Exp.Eye Res. 57: 763 - 769.
Goto Top

24. Wiechmann, A.F. and Hammarback, J.A. Molecular cloning and nucleotide sequence of a cDNA encoding recoverin from human retina. 1993; Exp.Eye Res. 56: 463 - 470.
Goto Top

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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