Angiotensin IV induces tyrosine phosphorylation of focal adhesion kinase and paxillin in proximal tubule cells

被引:34
作者
Chen, JK
Zimpelmann, J
Harris, RC
Burns, KD
机构
[1] Univ Ottawa, Ottawa Hosp Res Inst, Kidney Res Ctr, Dept Med,Div Nephrol, Ottawa, ON K1H 8L6, Canada
[2] Vanderbilt Univ, Dept Med, Nashville, TN 37232 USA
关键词
renin-angiotensin system; receptor; signaling; tyrosine kinase; renal proximal tubular epithelial cells;
D O I
10.1152/ajprenal.2001.280.6.F980
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
Angiotensin IV (ANG IV), the COOH-terminal hexapeptide fragment of angiotensin II (ANG II), binds to specific sites in the kidney, distinct from type 1 (AT(1)) and type 2 (AT(2)) receptors and designated type 4 (AT(4)) receptors. We determined signaling pathways for ANG IV in a proximal tubular cell line, LLC-PK(1)/Cl(4). In these cells, we found no specific binding of [(125)I]-ANG II. In contrast, ANG IV dose dependently competed for [(125)I]-labeled ANG IV binding, with no displacement by either ANG II, the AT1 receptor antagonist losartan, or the AT(2) antagonist PD-123319. Saturation binding indicated the presence of AT4 receptors of high affinity [dissociation constant (K(d)) = 1.4 nM]. ANG IV did not affect cAMP or cGMP production and did not increase cytosolic calcium concentration in these cells. In contrast, immunoprecipitation and immunoblotting studies revealed that ANG IV caused dose-dependent tyrosine phosphorylation of p125-focal adhesion kinase (p125-FAK) and p68-paxillin within 2 min, with maximal stimulation at 30 min. ANG IV-stimulated tyrosine phosphorylation of p125-FAK and paxillin was not affected by pretreatment with either losartan or PD-123319, and ANG II (10(-7) M) did not induce protein tyrosine phosphorylation. Our results indicate that LLC-PK(1)/Cl(4) cells express ANG IV receptors, which we demonstrate for the first time are linked to tyrosine phosphorylation of focal adhesion-associated proteins. This suggests that ANG IV, a product of ANG II metabolism, may regulate function of the focal adhesion complex in proximal tubule cells.
引用
收藏
页码:F980 / F988
页数:9
相关论文
共 52 条
[1]   DEVELOPMENT OF NA+-DEPENDENT HEXOSE-TRANSPORT IN A CULTURED LINE OF PORCINE KIDNEY-CELLS [J].
AMSLER, K ;
COOK, JS .
AMERICAN JOURNAL OF PHYSIOLOGY, 1982, 242 (01) :C94-C101
[2]   ANGIOTENSIN-II STIMULATION OF PROTEIN-SYNTHESIS AND CELL-GROWTH IN CHICK HEART-CELLS [J].
BAKER, KM ;
ACETO, JF .
AMERICAN JOURNAL OF PHYSIOLOGY, 1990, 259 (02) :H610-H618
[3]   Tyrosine kinase inhibition affects type 1 angiotensin II receptor internalization [J].
Becker, BN ;
Kondo, S ;
Chen, JK ;
Harris, RC .
JOURNAL OF RECEPTOR AND SIGNAL TRANSDUCTION RESEARCH, 1999, 19 (06) :975-993
[4]   Characterization of AT4 receptor from bovine aortic endothelium with photosensitive analogues of angiotensin IV [J].
Bernier, SG ;
Bellemare, JML ;
Escher, E ;
Guillemette, G .
BIOCHEMISTRY, 1998, 37 (12) :4280-4287
[5]   Study on the functionality and molecular properties of the AT4 receptor [J].
Briand, SI ;
Bellemare, JML ;
Bernier, SG ;
Guillemette, G .
ENDOCRINE RESEARCH, 1998, 24 (3-4) :315-323
[6]   Immortalized rabbit cortical collecting duct cells express AT(1) angiotensin II receptors [J].
Burns, KD ;
Regnier, L ;
Roczniak, A ;
Hebert, RL .
AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY, 1996, 271 (06) :F1147-F1157
[7]   SIGNALING AND GROWTH-RESPONSES OF LLC-PK1/CL-4 CELLS TRANSFECTED WITH THE RABBIT AT(1) ANG-II RECEPTOR [J].
BURNS, KD ;
HARRIS, RC .
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY, 1995, 268 (04) :C925-C935
[8]  
CARRY LA, 1999, FRONT BIOSCI, V4, pD102
[9]   Characterization of angiotensin IV-degrading enzymes and receptors on rat mesangial cells [J].
Chansel, D ;
Czekalski, S ;
Vandermeersch, S ;
Ruffet, E ;
Fournié-Zaluski, MC ;
Ardaillou, R .
AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY, 1998, 275 (04) :F535-F542
[10]   Transfection of an active cytochrome P450 arachidonic acid epoxygenase indicates that 14,15-epoxyeicosatrienoic acid functions as an intracellular second messenger in response to epidermal growth factor [J].
Chen, JK ;
Wang, DW ;
Falck, JR ;
Capdevila, J ;
Harris, RC .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (08) :4764-4769