Interaction of extracellular domain 2 of the human retina-specific ATP-binding cassette transporter (ABCA4) with all-trans-retinal.
The retina-specific ATP-binding cassette (ABC) transporter, ABCA4, is essential for transport of all-trans-retinal from the rod outer segment discs in the retina and is associated with a broad range of inherited retinal diseases, including Stargardt disease, autosomal recessive cone rod dystrophy, and fundus flavimaculatus. A unique feature of the ABCA subfamily of ABC transporters is the presence of highly conserved, long extracellular loops or domains (ECDs) with unknown function. The high degree of sequence conservation and mapped disease-associated mutations in these domains suggests an important physiological significance. Conformational analysis using CD spectroscopy of purified, recombinant ECD2 protein demonstrated that it has an ordered and stable structure composed of 27 +/- 3% alpha-helix, 20 +/- 3% beta-pleated sheet, and 53 +/- 3% coil. Significant conformational changes were observed in disease-associated mutant proteins. Using intrinsic tryptophan fluorescence emission spectrum of ECD2 polypeptide and fluorescence anisotropy, we have demonstrated that this domain specifically interacts with all-trans-retinal. Furthermore, the retinal interaction appeared preferential for the all-trans-isomer and was directly measurable through fluorescence anisotropy analysis. Our results demonstrate that the three macular degeneration-associated mutations lead to significant changes in the secondary structure of the ECD2 domain of ABCA4, as well as in its interaction with all-trans-retinal.
Biswas-Fiss
Esther E
EE
Department of Bioscience Technologies, Jefferson School of Health Professions, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA. esther.biswas@jefferson.edu
Kurpad
Deepa S
DS
Joshi
Kinjalben
K
Biswas
Subhasis B
SB
D018528:ATP-Binding Cassette Transporters / Q000737:chemistry*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D016880:Anisotropy; D002942:Circular Dichroism; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012160:Retina / Q000378:metabolism; D012172:Retinaldehyde / Q000737:chemistry*; D017386:Sequence Homology, Amino Acid; D014785:Vision, Ocular
C056475:ABCA4 protein, human; D018528:ATP-Binding Cassette Transporters; D000255:Adenosine Triphosphate; D012172:Retinaldehyde
BMC Struct Biol. 2008 May 13;8:25
18477405
Res Microbiol. 2005 Apr;156(3):322-7
15808935
Genomics. 1999 Sep 1;60(2):234-7
10486215
Hum Mol Genet. 2001 Nov 1;10(23):2671-8
11726554
Mol Ther. 2010 Mar;18(3):643-50
19953081
Biochim Biophys Acta. 2009 Jul;1791(7):573-83
19230850
EMBO J. 1996 Aug 1;15(15):3813-22
8670886
Physiol Rev. 1999 Jan;79(1 Suppl):S23-45
9922375
Eur J Biochem. 1993 Jan 15;211(1-2):267-80
8425537
Am J Hum Genet. 2000 Aug;67(2):487-91
10880298
Eur J Biochem. 2000 Sep;267(17):5306-12
10951189
Genet Mol Res. 2008 Feb 01;7(1):117-26
18273827
Invest Ophthalmol Vis Sci. 2004 Jun;45(6):1705-11
15161829
Mol Cell Endocrinol. 1993 Feb;91(1-2):35-41
8472852
Genome Res. 2001 Jul;11(7):1156-66
11435397
J Biol Chem. 1999 Mar 19;274(12):8269-81
10075733
Nucleic Acids Res. 2000 Jul 1;28(13):2494-502
10871398
J Mol Evol. 2005 Jan;60(1):72-80
15696369
Cancer Chemother Pharmacol. 1997;40 Suppl:S3-8
9272126
Biochemistry. 2000 May 9;39(18):5518-26
10820025
Mol Vis. 2009 Jul 28;15:1429-44
19641632
Biochemistry. 1995 May 23;34(20):6700-10
7756301
Nature. 1970 Aug 15;227(5259):680-5
5432063
Methods Enzymol. 2000;315:864-79
10736746
EMBO J. 1982;1(8):945-51
6329717
Biochemistry. 2009 Jul 21;48(28):6684-91
19507822
Anal Biochem. 1976 May 7;72:248-54
942051
Invest Ophthalmol Vis Sci. 2006 Dec;47(12):5137-52
17122096
J Mol Biol. 1982 May 5;157(1):105-32
7108955
Proc Natl Acad Sci U S A. 2004 Apr 20;101(16):5928-33
15067110
J Biol Chem. 1997 Nov 28;272(48):30405-11
9374531
J Biol Chem. 2001 Jun 29;276(26):23539-46
11320094
J Biol Chem. 2000 Jul 7;275(27):20399-405
10767284
Biophys Chem. 1988 Dec;32(2-3):173-85
3251567
J Biol Chem. 2002 Jun 14;277(24):21759-67
11919200
Biochemistry. 2001 Jul 27;40(28):8181-7
11444963
J Cell Biol. 1978 Aug;78(2):415-25
690173
Invest Ophthalmol Vis Sci. 2002 Jun;43(6):1980-5
12037008
J Biol Chem. 2005 Jan 14;280(2):1625-33
15528208
EMBO J. 1991 Dec;10(12):3777-85
1935899
Annu Rev Cell Biol. 1992;8:67-113
1282354
J Bioenerg Biomembr. 2007 Dec;39(5-6):507-17
17994272
Hum Mutat. 2003 Nov;22(5):395-403
14517951
Arch Ophthalmol. 2003 Jun;121(6):851-5
12796258
N Engl J Med. 2009 Dec 17;361(25):2483-4
20018970
J Biol Chem. 1997 Apr 11;272(15):10303-10
9092582
Science. 2001 Sep 7;293(5536):1782-4
11546861
Protein Eng. 2001 Jan;14(1):7-15
11287674
Graefes Arch Clin Exp Ophthalmol. 2005 Feb;243(2):90-100
15614537
Invest Ophthalmol Vis Sci. 2001 Sep;42(10):2229-36
11527935
Arch Ophthalmol. 1999 Apr;117(4):504-10
10206579
Protein Eng. 1993 Jun;6(4):383-90
8332596
Protein Expr Purif. 2007 Mar;52(1):40-9
16982201
Biochemistry. 2006 Mar 21;45(11):3813-23
16533065
Eur J Biochem. 2000 Jul;267(13):4242-52
10866829
Am J Hum Genet. 1999 Feb;64(2):422-34
9973280
J Biol Chem. 1994 Apr 8;269(14):10370-7
8144619
Res Microbiol. 2001 Apr-May;152(3-4):205-10
11421269
Biochemistry. 2003 Sep 16;42(36):10683-96
12962493
Arch Biochem Biophys. 2009 Dec;492(1-2):17-28
19818729
Nat Genet. 1997 Mar;15(3):236-46
9054934
J Biol Chem. 2004 Dec 24;279(52):53972-9
15471866
Biochemistry. 1999 Jun 8;38(23):7453-61
10360942
Invest Ophthalmol Vis Sci. 2001 May;42(6):1179-89
11328725
Invest Ophthalmol Vis Sci. 2000 Feb;41(2):337-48
10670460
Acta Ophthalmol Scand. 1998 Dec;76(6):649-52
9881544
J Immunol Methods. 2008 May 20;334(1-2):21-8
18346754
Cell. 1999 Jul 9;98(1):13-23
10412977
Methods Enzymol. 1998;292:213-24
9711556
Proc Natl Acad Sci U S A. 2007 Oct 30;104(44):17323-8
17947383