title: The role of PSD-95 in the rearrangement of Kv1.3 channels to the immunological synapse.
abstract: Establishment of the immunological synapse (IS) between T lymphocytes and antigen-presenting cells is a key step in the adaptive immune response. Several proteins accumulate in the IS, such as the Kv1.3 potassium channel; however, the mechanism of this translocation is unknown. PSD-95 and SAP97 are adaptor proteins that regulate the polarized cell surface expression and localization of Kv1 channels in neurons. We investigated whether these proteins affect the redistribution of Kv1.3 into the IS in non-excitable human T cells. We show here that PSD-95 and SAP97 are expressed in Jurkat and interact with the C terminus of Kv1.3. Disruption of the interaction between PSD-95 or SAP97 and Kv1.3 in Jurkat was realized by the expression of a C-terminal truncated Kv1.3, which lacks the binding domain for these proteins, or by the knockdown of the expression of PSD-95 or SAP97 using specific shRNA. Expression of the truncated Kv1.3 or knockdown of PSD-95, but not the knockdown of SAP97, inhibited the recruitment of Kv1.3 into the IS; the fraction of cells showing polarized Kv1.3 expression upon engagement in an IS was significantly lower than in control cells expressing the full-length Kv1.3, and the rearrangement of Kv1.3 did not show time dependence. In contrast, Jurkat cells expressing the full-length channel showed marked time dependence in the recruitment into the IS peaking at 1 min after the conjugation of the cells. These results demonstrate that PSD-95 participates in the targeting of Kv1.3 into the IS, implying its important role in human T-cell activation.
authors.affiliation: Department of Biophysics and Cell Biology, University of Debrecen, Nagyerdei krt. 98, 4032 Debrecen, Hungary.
authors.forename: Orsolya
authors.initials: O
authors.lastname: Szilágyi
authors.forename: Anita
authors.initials: A
authors.lastname: Boratkó
authors.forename: György
authors.initials: G
authors.lastname: Panyi
authors.forename: Péter
authors.initials: P
authors.lastname: Hajdu
mesh_terms: D001665:Binding Sites; D017353:Gene Deletion; D057809:HEK293 Cells; D006801:Humans; D054992:Immunological Synapses / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D019169:Jurkat Cells; D051664:Kv1.3 Potassium Channel / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D021381:Protein Transport
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D051664:Kv1.3 Potassium Channel
references.citation: Immunity. 2005 Jun;22(6):655-6
references.pmid: 15963778
references.citation: Neuropharmacology. 1996;35(7):993-1000
references.pmid: 8938729
references.citation: J Exp Med. 1993 Mar 1;177(3):637-45
references.pmid: 7679705
references.citation: J Biol Chem. 1999 Jul 16;274(29):20123-6
references.pmid: 10400624
references.citation: J Immunol. 2007 Jul 1;179(1):346-56
references.pmid: 17579055
references.citation: Annu Rev Immunol. 1997;15:707-47
references.pmid: 9143705
references.citation: Cell Death Differ. 2011 Jan;18(1):99-108
references.pmid: 20577261
references.citation: J Gen Physiol. 1996 Mar;107(3):409-20
references.pmid: 8868051
references.citation: Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10094-8
references.pmid: 1279670
references.citation: Science. 1997 Jan 3;275(5296):73-7
references.pmid: 8974395
references.citation: Curr Pharm Des. 2006;12(18):2199-220
references.pmid: 16787250
references.citation: J Biol Chem. 2000 Jan 14;275(2):1357-64
references.pmid: 10625685
references.citation: Nature. 1984 Feb 2-8;307(5950):468-71
references.pmid: 6320008
references.citation: Mol Membr Biol. 2007 May-Jun;24(3):194-205
references.pmid: 17520476
references.citation: Science. 1999 Sep 3;285(5433):1565-9
references.pmid: 10477520
references.citation: Proc Natl Acad Sci U S A. 2007 Sep 4;104(36):14418-23
references.pmid: 17726106
references.citation: Proc Natl Acad Sci U S A. 2007 Aug 7;104(32):13022-7
references.pmid: 17666528
references.citation: Immunol Rev. 2009 Sep;231(1):59-87
references.pmid: 19754890
references.citation: Am J Physiol Cell Physiol. 2012 May 15;302(10):C1504-12
references.pmid: 22378744
references.citation: EMBO J. 2011 Aug 16;30(19):3895-912
references.pmid: 21847095
references.citation: J Biol Chem. 1997 Oct 24;272(43):26899-904
references.pmid: 9341123
references.citation: J Theor Biol. 2012 May 7;300:173-82
references.pmid: 22285786
references.citation: Annu Rev Immunol. 2001;19:375-96
references.pmid: 11244041
references.citation: J Immunol. 2010 Jan 1;184(1):184-90
references.pmid: 19949095
references.citation: J Neurochem. 2007 Nov;103(4):1608-27
references.pmid: 17854350
references.citation: Nature. 1995 Nov 2;378(6552):85-8
references.pmid: 7477295
references.citation: Cell. 1999 Mar 5;96(5):611-4
references.pmid: 10089876
references.citation: Pflugers Arch. 1981 Aug;391(2):85-100
references.pmid: 6270629
references.citation: Biochem Biophys Res Commun. 2003 Jun 13;305(4):890-7
references.pmid: 12767914
references.citation: Proc Natl Acad Sci U S A. 2008 Feb 12;105(6):2011-6
references.pmid: 18250319
references.citation: J Biol Chem. 2001 Apr 13;276(15):12249-56
references.pmid: 11278890
references.citation: J Immunol. 2009 Nov 15;183(10):6296-302
references.pmid: 19841189
references.citation: Exp Cell Res. 2011 Oct 1;317(16):2333-41
references.pmid: 21726550
references.citation: Trends Pharmacol Sci. 2004 May;25(5):280-9
references.pmid: 15120495
references.citation: Cell Calcium. 2010 Jan;47(1):19-28
references.pmid: 19959227
references.citation: Am J Physiol Cell Physiol. 2007 Apr;292(4):C1431-9
references.pmid: 17151145
references.citation: Proc Natl Acad Sci U S A. 2006 Nov 14;103(46):17414-9
references.pmid: 17088564
references.citation: Proc Natl Acad Sci U S A. 2004 Feb 3;101(5):1285-90
references.pmid: 14745040
references.citation: Proc Natl Acad Sci U S A. 1997 Jul 8;94(14):7661-6
references.pmid: 9207149
references.citation: J Cell Biol. 2000 Jan 10;148(1):147-58
references.pmid: 10629225
references.citation: J Gen Physiol. 1999 Jan;113(1):71-80
references.pmid: 9874689
references.citation: Blood. 2010 Nov 4;116(18):3475-84
references.pmid: 20660288