Arginine-glycine-aspartic acid (RGD)-containing peptides inhibit the force production of mouse papillary muscle bundles via alpha 5 beta 1 integrin.
Integrins are considered to be an important mechanosensor in cardiac myocytes. To test whether integrins can influence cardiac contractile function, the force-frequency relationships of mouse papillary muscle bundles were measured in the presence or absence of a synthetic integrin-binding peptide, GRGDNP (gly-arg-gly-asp-asn-pro). Results demonstrate that in the presence of an arginine-glycine-aspartic acid (RGD)-containing synthetic peptide, contractile force was depressed significantly by, 28% at 4 Hz, 37.7% at 5 Hz and 20% at 10 Hz (n = 6, P < 0.01). Treatment of myofibres with either protease-generated fragments of denatured collagen (Type I) or denatured collagen that contain the RGD motif, also reduced force production significantly. An integrin-activating antibody for beta(1) integrin inhibited the force similar to synthetic RGD peptide. Function-blocking integrin antibodies for alpha(5) and beta(1) integrins reversed the effect of the RGD-containing peptide, and alpha(5) integrin also reversed the effect of proteolytic fragments of denatured collagen on contractile force, whereas experiments with function-blocking antibody for beta(3) integrin did not reverse the effect of RGD peptide. Force-[Ca(2)(+)](i) measurements showed that the depressed rate of force generation observed in the presence of the RGD-containing peptide was associated with reduced [Ca(2)(+)](i). Data analyses further demonstrated that force per unit of Ca(2)(+) was reduced, suggesting that the myofilament activation process was altered. In addition, inhibition of PKC enzyme using the selective, cell-permeable inhibitor Ro-32-0432, reversed the activity of RGD peptide on papillary muscle bundles. In conclusion, these data indicate that RGD peptide, acting via alpha(5)beta(1) integrin, depresses the force production from papillary muscle bundles, partly associated with changes in [Ca(2)(+)](i) and the myofilament activation processes, that is modulated by PKCepsilon.
Sarin
Vandana
V
Department of Medical Physiology, Cardiovascular Research Institute, 336 Reynolds Medical Building, Texas A & M University System Health Science Center, College of Medicine, College Station, TX 77843-1114, USA.
Gaffin
Robert D
RD
Meininger
Gerald A
GA
Muthuchamy
Mariappan
M
D000818:Animals; D004305:Dose-Response Relationship, Drug; D066298:In Vitro Techniques; D039081:Integrin alpha5beta1 / Q000096:biosynthesis* / Q000235:genetics; D008297:Male; D051379:Mice; D009119:Muscle Contraction / Q000187:drug effects / Q000502:physiology*; D009842:Oligopeptides / Q000494:pharmacology*; D010210:Papillary Muscles / Q000187:drug effects* / Q000378:metabolism
D039081:Integrin alpha5beta1; D009842:Oligopeptides; C047981:arginyl-glycyl-aspartic acid
J Physiol. 2004 Dec 15;561(Pt 3):777-91
15486021
J Natl Cancer Inst. 1956 Jun;16(6):1375-403
13320119
Cardiovasc Res. 1999 Aug 1;43(2):371-81
10536667
Exp Cell Res. 2000 Jan 10;254(1):55-63
10623465
Am J Physiol Heart Circ Physiol. 2000 Mar;278(3):H769-79
10710345
Am J Cardiol. 1999 Jun 17;83(12A):38H-43H
10750585
Am J Pathol. 2000 May;156(5):1489-98
10793060
Circ Res. 2000 Sep 15;87(6):508-15
10988244
Am J Physiol Heart Circ Physiol. 2001 Jan;280(1):H361-7
11123252
J Mol Cell Cardiol. 2001 Feb;33(2):271-82
11162132
Circ Res. 2001 Jun 8;88(11):1112-9
11397776
Cell Tissue Res. 2001 Sep;305(3):285-98
11572082
Circ Res. 2002 Mar 8;90(4):458-64
11884376
Circ Res. 2002 Mar 8;90(4):473-80
11884378
Trends Cardiovasc Med. 2002 Apr;12(3):97-101
12007733
Am J Physiol Heart Circ Physiol. 2003 Jan;284(1):H122-32
12485818
Circulation. 2003 Feb 25;107(7):1046-52
12600920
Cardiovasc Res. 2003 Mar;57(3):766-74
12618238
J Biol Chem. 2003 Mar 28;278(13):11265-72
12551921
J Vasc Res. 2003 May-Jun;40(3):211-33
12902635
J Biol Chem. 2003 Oct 24;278(43):42214-24
12909616
Am J Physiol Cell Physiol. 2004 Mar;286(3):C518-28
14761883
Am J Physiol Heart Circ Physiol. 2004 Mar;286(3):H889-94
14576082
J Biol Chem. 1985 Mar 25;260(6):3440-50
3838314
Biochem Biophys Res Commun. 1992 Feb 14;182(3):1025-31
1540151
Cell Motil Cytoskeleton. 1992;21(2):87-100
1373110
Proc Natl Acad Sci U S A. 1993 Oct 1;90(19):9051-5
7692444
Biophys J. 1993 Aug;65(2):693-701
8218897
Biophys J. 1994 Jul;67(1):273-82
7918995
Cardiovasc Drugs Ther. 1995 Aug;9(4):581-7
8547208
Biochemistry. 2004 May 25;43(20):5996-6004
15147183
Mol Biol Cell. 2004 Jun;15(6):2558-67
15034138
Cardiovasc Res. 2004 Aug 15;63(3):381-90
15276463
J Physiol. 2004 Aug 1;558(Pt 3):927-41
15194741
Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4
388439
Circ Res. 1996 Oct;79(4):821-6
8831506
Biophys J. 1996 Oct;71(4):1891-904
8889164
J Mol Biol. 1997 Feb 14;266(1):8-14
9054965
Am J Physiol. 1997 Jul;273(1 Pt 2):H175-82
9249488
J Clin Invest. 1997 Sep 15;100(6):1647-53
9294134
J Biol Chem. 1998 Mar 20;273(12):6670-8
9506964
J Cell Sci. 1998 Apr;111 ( Pt 8):1117-26
9512507
Circ Res. 1998 Sep 7;83(5):471-80
9734469
J Cell Biol. 1998 Oct 5;143(1):241-52
9763435
Circ Res. 1999 Jan 8-22;84(1):122-4
9915781
Am J Physiol. 1999 Feb;276(2 Pt 2):H642-50
9950866
Lab Invest. 1958 Mar-Apr;7(2):134-7
13540204
J Biol Chem. 1996 Oct 11;271(41):25099-106
8810264