title: Bradykinin B2 receptor interacts with integrin alpha5beta1 to transactivate epidermal growth factor receptor in kidney cells.
abstract: We have shown previously that the vasoactive peptide bradykinin (BK) stimulates proliferation of a cultured murine cell model of the inner medullary collecting duct (mIMCD-3 cells) via transactivation of epidermal growth factor receptor (EGFR) by a mechanism that involves matrix metalloproteinases (collagenase-2 and -3). Because collagenases lack an integral membrane domain, we hypothesized that receptors for extracellular matrix proteins, integrins, may play a role in BK-induced signaling by targeting collagenases to the membrane, thus forming a functional signaling complex. BK-induced phosphorylation of extracellular signal-regulated protein kinase (ERK) in mIMCD-3 cells was reduced by approximately 65% by synthetic peptides containing an Arg-Gly-Asp sequence, supporting roles for integrins in BK-induced signaling. Neutralizing antibody against alpha5beta1 integrin partially (approximately 60%) blocked BK-induced ERK activation but did not affect EGF-induced ERK activation. Silencing of alpha5 and beta1 expression by transfecting cells with small interfering RNAs (siRNA) significantly decreased BK-induced ERK activation (approximately 80%) and EGFR phosphorylation (approximately 50%). This effect was even more pronounced in cells that were cotransfected with siRNAs directed against both collagenases and alpha5beta1 integrin. On the basis of our results, we suggested that integrin alpha5beta1 is involved in BK-induced signaling in mIMCD-3 cells. Using immunoprecipitation/Western blotting, we demonstrated association of BK B(2) receptor with alpha5beta1 integrin upon BK treatment. Furthermore, BK induced association of alpha5beta1 integrin with EGFR. These data provide the first evidence that specific integrins are involved in BK B(2) receptor-induced signaling in kidney cells, and ultimately might lead to development of new strategies for treatment of renal tubulointerstitial fibrosis.
authors.affiliation: Medical and Research Services, Ralph H. Johnson Veterans Affairs Medical Center, Medical University of South Carolina, Charleston, SC 29425-6290, USA.
authors.forename: Inga I
authors.initials: II
authors.lastname: Kramarenko
authors.forename: Marlene A
authors.initials: MA
authors.lastname: Bunni
authors.forename: John R
authors.initials: JR
authors.lastname: Raymond
authors.forename: Maria N
authors.initials: MN
authors.lastname: Garnovskaya
mesh_terms: D000818:Animals; D002460:Cell Line; D004789:Enzyme Activation; D066246:ErbB Receptors / Q000235:genetics*; D039081:Integrin alpha5beta1 / Q000378:metabolism*; D007668:Kidney / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D020782:Matrix Metalloproteinases / Q000235:genetics; D051379:Mice; D010766:Phosphorylation; D011485:Protein Binding; D034741:RNA, Small Interfering; D043782:Receptor, Bradykinin B2 / Q000378:metabolism*; D015533:Transcriptional Activation
chemical_list: D039081:Integrin alpha5beta1; D034741:RNA, Small Interfering; D043782:Receptor, Bradykinin B2; D066246:ErbB Receptors; D020782:Matrix Metalloproteinases
references.citation: Nature. 1999 Dec 23-30;402(6764):884-8
references.pmid: 10622253
references.citation: J Clin Invest. 1985 Jul;76(1):132-6
references.pmid: 4019771
references.citation: Biochem Pharmacol. 2009 Jan 15;77(2):269-76
references.pmid: 18938142
references.citation: J Biol Chem. 2005 Nov 25;280(47):39050-7
references.pmid: 16186116
references.citation: J Biol Chem. 1999 May 14;274(20):13978-84
references.pmid: 10318809
references.citation: Am J Physiol. 1990 Jun;258(6 Pt 2):F1584-91
references.pmid: 2163214
references.citation: J Pharmacol Exp Ther. 2003 Mar;304(3):968-77
references.pmid: 12604671
references.citation: Biochem Soc Trans. 2004 Jun;32(Pt3):438-42
references.pmid: 15157155
references.citation: J Pharmacol Exp Ther. 2006 Sep;318(3):1033-43
references.pmid: 16717107
references.citation: Annu Rev Pharmacol Toxicol. 2002;42:283-323
references.pmid: 11807174
references.citation: Proc Natl Acad Sci U S A. 1998 Jun 23;95(13):7281-6
references.pmid: 9636140
references.citation: J Cell Biol. 2001 Apr 30;153(3):491-501
references.pmid: 11331301
references.citation: J Biol Chem. 2000 Oct 20;275(42):32736-46
references.pmid: 10915788
references.citation: J Biol Chem. 2000 Feb 25;275(8):5291-6
references.pmid: 10681501
references.citation: J Biol Chem. 2001 Aug 3;276(31):29368-74
references.pmid: 11359786
references.citation: J Biol Chem. 2003 Sep 5;278(36):34674-84
references.pmid: 12824186
references.citation: J Biol Chem. 2000 Jul 21;275(29):21785-8
references.pmid: 10801897
references.citation: J Biol Chem. 2001 Aug 3;276(31):29375-81
references.pmid: 11359774
references.citation: J Biol Chem. 2004 Jan 16;279(3):1980-93
references.pmid: 14559894
references.citation: Blood. 2006 Sep 1;108(5):1441-50
references.pmid: 16609063
references.citation: Am J Physiol. 1996 Aug;271(2 Pt 2):R352-60
references.pmid: 8770134
references.citation: J Cell Sci. 2006 Oct 1;119(Pt 19):3901-3
references.pmid: 16988024
references.citation: Neuroscience. 2007 Feb 9;144(3):889-97
references.pmid: 17157995
references.citation: Cell. 1998 Feb 6;92(3):391-400
references.pmid: 9476898
references.citation: J Biol Chem. 2000 Aug 11;275(32):24907-14
references.pmid: 10801799
references.citation: Nat Cell Biol. 2002 Apr;4(4):E65-8
references.pmid: 11944032
references.citation: J Biol Chem. 2000 Apr 28;275(17):12970-7
references.pmid: 10777598
references.citation: Cell. 1996 May 31;85(5):683-93
references.pmid: 8646777
references.citation: J Neurochem. 2005 Nov;95(3):630-40
references.pmid: 16135088
references.citation: Proc Natl Acad Sci U S A. 2003 Aug 5;100(16):9482-7
references.pmid: 12874388
references.citation: EMBO J. 1998 Nov 16;17(22):6622-32
references.pmid: 9822606
references.citation: Cell Signal. 2008 Dec;20(12):2324-31
references.pmid: 18804531
references.citation: J Biol Chem. 2003 Jun 20;278(25):22631-43
references.pmid: 12692126
references.citation: J Biol Chem. 2004 Feb 27;279(9):8056-62
references.pmid: 14679206
references.citation: Nat Cell Biol. 2002 Apr;4(4):E83-90
references.pmid: 11944041