Identification of the retinoschisin-binding site on the retinal Na/K-ATPase.
Adenosine Triphosphatases
/ genetics
Animals
Binding Sites
Cation Transport Proteins
/ genetics
Cell Adhesion Molecules
/ genetics
Cell Adhesion Molecules, Neuronal
/ genetics
Eye Proteins
/ genetics
HEK293 Cells
Humans
Mice
Mice, Knockout
Mutagenesis, Site-Directed
Retina
/ metabolism
Sodium-Potassium-Exchanging ATPase
/ genetics
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2019
2019
Historique:
received:
11
09
2018
accepted:
19
04
2019
entrez:
4
5
2019
pubmed:
3
5
2019
medline:
15
1
2020
Statut:
epublish
Résumé
X-linked juvenile retinoschisis (XLRS) is a hereditary retinal dystrophy, caused by mutations in the RS1 gene which encodes the secreted protein retinoschisin. In recent years, several molecules have been proposed to interact with retinoschisin, including the retinal Na/K-ATPase, L-voltage gated Ca2+ channels, and specific sugars. We recently showed that the retinal Na/K-ATPase consisting of subunits ATP1A3 and ATP1B2 is essential for anchoring retinoschisin to plasma membranes and identified the glycosylated ATP1B2 subunit as the direct interaction partner for retinoschisin. We now aimed to precisely map the retinoschisin binding domain(s) in ATP1B2. In general, retinoschisin binding was not affected after selective elimination of individual glycosylation sites via site-directed mutagenesis as well as after full enzymatic deglycosylation of ATP1B2. Applying the interface prediction tool PresCont, two putative protein-protein interaction patches ("patch I" and "patch II") consisting each of four hydrophobic amino acid stretches on the ATP1B2 surface were identified. These were consecutively altered by site-directed mutagenesis. Functional assays with the ATP1B2 patch mutants identified patch II and, specifically, the associated amino acid at position 240 (harboring a threonine in ATP1B2) as crucial for retinoschisin binding to ATP1B2. These and previous results led us to suggest an induced-fit binding mechanism for the interaction between retinoschisin and the Na/K-ATPase, which is dependent on threonine 240 in ATP1B2 allowing the accommodation of hyperflexible retinoschisin spikes by the associated protein-protein interaction patch on ATP1B2.
Identifiants
pubmed: 31048931
doi: 10.1371/journal.pone.0216320
pii: PONE-D-18-26568
pmc: PMC6497308
doi:
Substances chimiques
Atp1b2 protein, mouse
0
Cation Transport Proteins
0
Cell Adhesion Molecules
0
Cell Adhesion Molecules, Neuronal
0
Eye Proteins
0
RS1 protein, mouse
0
Adenosine Triphosphatases
EC 3.6.1.-
Atp1a3 protein, mouse
EC 3.6.1.-
Sodium-Potassium-Exchanging ATPase
EC 7.2.2.13
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0216320Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
Références
Biochemistry. 2010 Aug 24;49(33):7023-32
pubmed: 20677810
Proteins. 2000 Jun 1;39(4):331-42
pubmed: 10813815
Mol Vis. 2006 Aug 10;12:892-901
pubmed: 16917482
Hum Mol Genet. 2011 Mar 15;20(6):1132-42
pubmed: 21196491
Mol Vis. 2006 Sep 28;12:1108-16
pubmed: 17093404
Proteins. 2012 Jan;80(1):154-68
pubmed: 22038731
J Cell Biol. 1987 Jun;104(6):1587-95
pubmed: 2438288
Biochemistry. 2008 Sep 2;47(35):9098-106
pubmed: 18690710
Biochim Biophys Acta. 2007 Sep;1774(9):1069-78
pubmed: 17702679
Annu Rev Biochem. 2002;71:511-35
pubmed: 12045105
Sci Rep. 2014 Jun 04;4:5165
pubmed: 24893715
Virology. 2000 Aug 15;274(1):75-85
pubmed: 10936090
Ann N Y Acad Sci. 1974;242(0):598-616
pubmed: 4279608
Immunobiology. 2010 Sep-Oct;215(9-10):685-91
pubmed: 20573418
Protein Sci. 1998 Jul;7(7):1626-31
pubmed: 9684896
Vision Res. 2001 Dec;41(28):3931-42
pubmed: 11738458
Biochemistry. 2018 Feb 20;57(7):1249-1261
pubmed: 29345906
Adv Pharm Bull. 2017 Apr;7(1):141-150
pubmed: 28507948
Proc Natl Acad Sci U S A. 2002 Apr 30;99(9):6222-7
pubmed: 11983912
Cell Mol Life Sci. 2013 Jan;70(2):205-22
pubmed: 22695678
J Biochem. 2019 Jun 1;165(6):487-495
pubmed: 30597085
Electrophoresis. 2016 Jun;37(11):1407-19
pubmed: 26872045
Adv Exp Med Biol. 2002;514:439-50
pubmed: 12596937
Exp Eye Res. 2018 Dec;177:23-34
pubmed: 30040949
J Mol Biol. 1987 Aug 5;196(3):641-56
pubmed: 3681970
Proc Natl Acad Sci U S A. 2003 May 13;100(10):5772-7
pubmed: 12730379
Methods Mol Biol. 2016;1377:305-18
pubmed: 26695042
Invest Ophthalmol Vis Sci. 2007 Mar;48(3):991-1000
pubmed: 17325137
BMC Bioinformatics. 2008 Jan 23;9:40
pubmed: 18215316
J Mol Biol. 2004 Jun 25;340(1):81-95
pubmed: 15184024
Proc Natl Acad Sci U S A. 2016 May 10;113(19):5287-92
pubmed: 27114531
J Biol Chem. 2009 Feb 6;284(6):3966-75
pubmed: 19074145
Anal Biochem. 1989 Aug 1;180(2):195-204
pubmed: 2510544
Mol Biol Cell. 2017 Aug 1;28(16):2178-2189
pubmed: 28615319
J Biol Chem. 2010 Dec 10;285(50):39289-302
pubmed: 20937802
J Biol Chem. 2007 Nov 9;282(45):32792-801
pubmed: 17804407
J Biol Chem. 2000 Jan 21;275(3):1976-86
pubmed: 10636900
Eur J Pharmacol. 1998 Sep 11;357(1):73-82
pubmed: 9788776
Front Cell Neurosci. 2017 Aug 08;11:232
pubmed: 28848397
Nat Rev Mol Cell Biol. 2004 Apr;5(4):282-95
pubmed: 15071553
J Cell Mol Med. 2017 Apr;21(4):768-780
pubmed: 27995734
Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12952-6
pubmed: 7809153
Surv Ophthalmol. 2004 Mar-Apr;49(2):214-30
pubmed: 14998693
Invest Ophthalmol Vis Sci. 2001 Mar;42(3):816-25
pubmed: 11222545
Hum Mol Genet. 2016 Dec 15;25(24):5311-5320
pubmed: 27798099
Biochemistry. 2010 Apr 13;49(14):3116-28
pubmed: 20199105
Nat Genet. 1997 Oct;17(2):164-70
pubmed: 9326935