THROMBIN RECEPTOR PEPTIDE INHIBITS THROMBIN-INDUCED INCREASE IN ENDOTHELIAL PERMEABILITY BY RECEPTOR DESENSITIZATION

被引:80
作者
LUM, H
ANDERSEN, TT
SIFLINGERBIRNBOIM, A
TIRUPPATHI, C
GOLIGORSKY, MS
FENTON, JW
MALIK, AB
机构
[1] ALBANY MED COLL,DEPT BIOCHEM & MOLEC BIOL,ALBANY,NY 12208
[2] NEW YORK STATE DEPT HLTH,WADSWORTH CTR LABS & RES,ALBANY,NY 12208
[3] SUNY STONY BROOK,DEPT MED,STONY BROOK,NY 11794
关键词
D O I
10.1083/jcb.120.6.1491
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Thrombin, a potent activator of cellular responses, proteolytically cleaves, and thereby activates its receptor. In the present study, we compared the effects of the thrombin receptor 14-amino acid peptide (TRP-14; SFLLRNPNDKYEPF), which comprises the NH2 terminus after cleavage of the thrombin receptor, and of the native alpha-thrombin on endothelial monolayer permeability. Addition of TRP-14 (1-200 muM) to bovine pulmonary artery endothelial cells increased [Ca2+]i, in a dose-dependent manner. The peak increase in [Ca2+]i in response to 100 muM TRP-14 or 0.1 muM alpha-thrombin was similar (i.e., 931 +/- 74 nM and 1032 +/- 80 nM, respectively), which was followed by a slow decrease with t1/2 values of 0.73 and 0.61 min, respectively. Extracellular Ca2+ chelation with 5 mM EGTA abolished the sustained increases in [Ca2+]i induced by either TRP-14 or alpha-thrombin. Alpha-thrombin (0.1 muM) increased transendothelial [I-125]albumin permeability, whereas TRP-14 (1-100 muM) had no effect. Coincubation of 100 muM TRP-14 with 1 muM DIP-alpha-thrombin also did not increase permeability over control values. Stimulation of BPAEC with 0.1 muM alpha-thrombin induced translocation of protein kinase C (PKC) from the cytosol to the plasma membrane indicative of PKC activation, whereas TRP-14 had no effect at any concentration. TRP-14 at 100 muM desensitized BPAEC to thrombin-induced increases in [Ca2+]i and transendothelial permeability. The Ca2+ desensitization was reversed after approximately 60 min, and this recovery paralleled the recovery of the permeability response. These findings indicate that the TRP-14-induced Ca2+ mobilization in the absence of PKC activation is insufficient to increase endothelial permeability. In contrast, the increase in endothelial permeability after alpha-thrombin occurred in conjunction with Ca2+ mobilization as well as PKC activation. TRP-14 pretreatment prevented the alpha-thrombin-induced increase in endothelial permeability secondary to desensitization of the Ca2+ signal. The results suggest that combined cytosolic Ca2+ mobilization mediated by TRP-14 and PKC activation mediated by a TRP-14-independent pathway are dual signals responsible for the thrombin-induced increase in vascular endothelial permeability.
引用
收藏
页码:1491 / 1499
页数:9
相关论文
共 36 条
  • [11] FENTON JW, 1988, ADV CLIN ENZYMOL, V6, P186
  • [12] THROMBIN-INDUCED INCREASE IN ALBUMIN PERMEABILITY ACROSS THE ENDOTHELIUM
    GARCIA, JGN
    SIFLINGERBIRNBOIM, A
    BIZIOS, R
    DELVECCHIO, PJ
    FENTON, JW
    MALIK, AB
    [J]. JOURNAL OF CELLULAR PHYSIOLOGY, 1986, 128 (01) : 96 - 104
  • [13] GOLIGORSKY MS, 1989, J BIOL CHEM, V264, P16771
  • [14] GRYNKIEWICZ G, 1985, J BIOL CHEM, V260, P3440
  • [15] HO T, 1988, J CELL BIOL, V107, P929
  • [16] ALPHA-THROMBIN-INDUCED PULMONARY VASOCONSTRICTION
    HORGAN, MJ
    FENTON, JW
    MALIK, AB
    [J]. JOURNAL OF APPLIED PHYSIOLOGY, 1987, 63 (05) : 1993 - 2000
  • [17] HUANG RS, 1991, J BIOL CHEM, V266, P18435
  • [18] JAFFE EA, 1987, J BIOL CHEM, V262, P8557
  • [19] COLOR TEST FOR DETECTION OF FREE TERMINAL AMINO GROUPS IN SOLID-PHASE SYNTHESIS OF PEPTIDES
    KAISER, E
    COLESCOT.RL
    BOSSINGE.CD
    COOK, PI
    [J]. ANALYTICAL BIOCHEMISTRY, 1970, 34 (02) : 595 - &
  • [20] HUMAN ALPHA-THROMBIN INDUCES PHOSPHOINOSITIDE TURNOVER AND CA-2+ MOVEMENTS IN CULTURED HUMAN UMBILICAL VEIN ENDOTHELIAL-CELLS
    LAMPUGNANI, MG
    PEDENOVI, M
    DEJANA, E
    ROTILIO, D
    DONATI, MB
    BUSSOLINO, F
    GARBARINO, G
    GHIGO, D
    BOSIA, A
    [J]. THROMBOSIS RESEARCH, 1989, 54 (01) : 75 - 87