Identification of a novel integrin α6β1 binding site in the angiogenic inducer CCN1 (CYR61)

被引:118
作者
Leu, SJ
Liu, Y
Chen, NY
Chen, CC
Lam, SCT
Lau, LF
机构
[1] Univ Illinois, Coll Med, Dept Mol Genet, Chicago, IL 60607 USA
[2] Univ Illinois, Coll Med, Dept Pharmacol, Chicago, IL 60607 USA
关键词
D O I
10.1074/jbc.M305862200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The angiogenic inducer CCN1 (cysteine-rich 61, CYR61), a secreted matricellular protein of the CCN family, is a ligand of multiple integrins, including alpha(6)beta(1). Previous studies have shown that CCN1 interaction with integrin alpha(6)beta(1) mediates adhesion of fibroblasts, endothelial cells, and smooth muscle cells, as well as migration of smooth muscle cells. Recently, we have reported that CCN1-induced tubule formation of unactivated endothelial cells is also mediated through integrin alpha(6)beta(1). In this study, we demonstrate that human skin fibroblasts adhere specifically to the T1 sequence (GQKCIVQTTSWSQCSKS) within domain III of CCN1, and this process is blocked by anti-alpha(6) and anti-beta(1) monoclonal antibodies. Alanine substitution mutagenesis of the T1 sequence further defines the sequence TTSWSQC-SKS as the critical determinant for mediating alpha(6)beta(1)-dependent adhesion. Soluble T1 peptide specifically inhibits fibroblast adhesion to CCN1 in a dose-dependent manner. Furthermore, T1 also inhibits cell adhesion to other alpha(6)beta(1) ligands, including CCN2 ( CTGF), CCN3 (NOV), and laminin, but not to ligands of other integrins. In addition, T1 specifically inhibits alpha(6)beta(1)-dependent tubule formation of unactivated endothelial cells in a CCN1-containing collagen gel matrix. To confirm that T1 binds integrin alpha(6)beta(1) directly, we perform affinity chromatography and show that integrin alpha(6)beta(1) is isolated from an octylglucoside extract of fibroblasts on T1- coupled Affi-gel. Taken together, these findings define the T1 sequence in CCN1 as a novel binding motif for integrin alpha(6)beta(1), providing the basis for the development of peptide mimetics to examine the functional role of alpha(6)beta(1) in angiogenesis.
引用
收藏
页码:33801 / 33808
页数:8
相关论文
共 48 条
  • [21] Adhesion of human umbilical vein endothelial cells to the immediate-early gene product Cyr61 is mediated through integrin αvβ3
    Kireeva, ML
    Lam, SCT
    Lau, LF
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (05) : 3090 - 3096
  • [22] HUMAN MICROVASCULAR ENDOTHELIAL-CELLS USE BETA-1 AND BETA-3 INTEGRIN RECEPTOR COMPLEXES TO ATTACH TO LAMININ
    KRAMER, RH
    CHENG, YF
    CLYMAN, R
    [J]. JOURNAL OF CELL BIOLOGY, 1990, 111 (03) : 1233 - 1243
  • [23] PROMOTER FUNCTION AND STRUCTURE OF THE GROWTH FACTOR-INDUCIBLE IMMEDIATE EARLY GENE CYR61
    LATINKIC, BV
    OBRIEN, TP
    LAU, LF
    [J]. NUCLEIC ACIDS RESEARCH, 1991, 19 (12) : 3261 - 3267
  • [24] The CCN family of angiogenic regulators: The integrin connection
    Lau, LF
    Lam, SCT
    [J]. EXPERIMENTAL CELL RESEARCH, 1999, 248 (01) : 44 - 57
  • [25] Pro-angiogenic activities of CYR61 (CCN1) mediated through integrins αvβ3 and α6β1 in human umbilical vein endothelial cells
    Leu, SJ
    Lam, SCT
    Lau, LF
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (48) : 46248 - 46255
  • [26] CCN3 (NOV) is a novel angiogenic regulator of the CCN protein family
    Lin, CG
    Leu, SJ
    Chen, NY
    Tebeau, CM
    Lin, SX
    Yeung, CY
    Lau, LF
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (26) : 24200 - 24208
  • [27] Two RGD-independent αvβ3 integrin binding sites on tumstatin regulate distinct anti-tumor properties
    Maeshima, Y
    Colorado, PC
    Kalluri, R
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (31) : 23745 - 23750
  • [28] CD36 associates with CD9 and integrins on human blood platelets
    Miao, WM
    Vasile, E
    Lane, WS
    Lawler, J
    [J]. BLOOD, 2001, 97 (06) : 1689 - 1696
  • [29] CYR61 (CCN1) is essential for placental development and vascular integrity
    Mo, FE
    Muntean, AG
    Chen, CC
    Stolz, DB
    Watkins, SC
    Lau, LF
    [J]. MOLECULAR AND CELLULAR BIOLOGY, 2002, 22 (24) : 8709 - 8720
  • [30] Murayama O, 1996, J BIOCHEM-TOKYO, V120, P445