Bradykinin B2 receptor interacts with integrin alpha5beta1 to transactivate epidermal growth factor receptor in kidney cells.
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.
Kramarenko
Inga I
II
Medical and Research Services, Ralph H. Johnson Veterans Affairs Medical Center, Medical University of South Carolina, Charleston, SC 29425-6290, USA.
Bunni
Marlene A
MA
Raymond
John R
JR
Garnovskaya
Maria N
MN
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
D039081:Integrin alpha5beta1; D034741:RNA, Small Interfering; D043782:Receptor, Bradykinin B2; D066246:ErbB Receptors; D020782:Matrix Metalloproteinases
Nature. 1999 Dec 23-30;402(6764):884-8
10622253
J Clin Invest. 1985 Jul;76(1):132-6
4019771
Biochem Pharmacol. 2009 Jan 15;77(2):269-76
18938142
J Biol Chem. 2005 Nov 25;280(47):39050-7
16186116
J Biol Chem. 1999 May 14;274(20):13978-84
10318809
Am J Physiol. 1990 Jun;258(6 Pt 2):F1584-91
2163214
J Pharmacol Exp Ther. 2003 Mar;304(3):968-77
12604671
Biochem Soc Trans. 2004 Jun;32(Pt3):438-42
15157155
J Pharmacol Exp Ther. 2006 Sep;318(3):1033-43
16717107
Annu Rev Pharmacol Toxicol. 2002;42:283-323
11807174
Proc Natl Acad Sci U S A. 1998 Jun 23;95(13):7281-6
9636140
J Cell Biol. 2001 Apr 30;153(3):491-501
11331301
J Biol Chem. 2000 Oct 20;275(42):32736-46
10915788
J Biol Chem. 2000 Feb 25;275(8):5291-6
10681501
J Biol Chem. 2001 Aug 3;276(31):29368-74
11359786
J Biol Chem. 2003 Sep 5;278(36):34674-84
12824186
J Biol Chem. 2000 Jul 21;275(29):21785-8
10801897
J Biol Chem. 2001 Aug 3;276(31):29375-81
11359774
J Biol Chem. 2004 Jan 16;279(3):1980-93
14559894
Blood. 2006 Sep 1;108(5):1441-50
16609063
Am J Physiol. 1996 Aug;271(2 Pt 2):R352-60
8770134
J Cell Sci. 2006 Oct 1;119(Pt 19):3901-3
16988024
Neuroscience. 2007 Feb 9;144(3):889-97
17157995
Cell. 1998 Feb 6;92(3):391-400
9476898
J Biol Chem. 2000 Aug 11;275(32):24907-14
10801799
Nat Cell Biol. 2002 Apr;4(4):E65-8
11944032
J Biol Chem. 2000 Apr 28;275(17):12970-7
10777598
Cell. 1996 May 31;85(5):683-93
8646777
J Neurochem. 2005 Nov;95(3):630-40
16135088
Proc Natl Acad Sci U S A. 2003 Aug 5;100(16):9482-7
12874388
EMBO J. 1998 Nov 16;17(22):6622-32
9822606
Cell Signal. 2008 Dec;20(12):2324-31
18804531
J Biol Chem. 2003 Jun 20;278(25):22631-43
12692126
J Biol Chem. 2004 Feb 27;279(9):8056-62
14679206
Nat Cell Biol. 2002 Apr;4(4):E83-90
11944041