EFFECT OF THE CONFORMATION AND ORIENTATION OF ADSORBED FIBRONECTIN ON ENDOTHELIAL-CELL SPREADING AND THE STRENGTH OF ADHESION
EFFECT OF THE CONFORMATION AND ORIENTATION OF ADSORBED FIBRONECTIN ON ENDOTHELIAL-CELL SPREADING AND THE STRENGTH OF ADHESION
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DOI：
10.1002/jbm.820270816
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发表时间：
1993-08-01
期刊：
影响因子：
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通讯作者：
TRUSKEY, GA
中科院分区：
文献类型：
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作者：
IULIANO, DJ;SAAVEDRA, SS;TRUSKEY, GA
研究了表面疏水性对纤维连接蛋白（Fn）细胞结合区构象的影响，以及Fn构象对牛主动脉内皮细胞（BAEC）粘附的影响。用丙烯酰二氢呋喃选择性标记Fn的游离巯基。在溶液中以及在吸附到亲水性玻璃和疏水性硅烷化玻璃时监测荧光发射。吸附后，丙烯酰二氢呋喃标记的Fn发射最大值向较长波长偏移，并且吸附到亲水性玻璃上的丙烯酰二氢呋喃标记的Fn的偏移大于吸附到疏水性硅烷上的丙烯酰二氢呋喃标记的Fn的偏移，这表明丙烯酰二氢呋喃在玻璃上比硅烷更易接近溶剂的环境中。BAEC，悬浮在无血清培养基中，附着15或120分钟到含有预吸附Fn的玻璃或硅烷表面上，之后，在平行板流动室中测量细胞铺展和粘附强度。细胞扩散是相似的两个表面后，15分钟的附件，但BAECs在玻璃上比硅烷120分钟后蔓延。在低表面浓度的Fn，BAECs更粘附在玻璃上比硅烷。在较高的表面浓度，粘附力是相似的。在无血清培养基中孵育2小时后，通过黏着斑蛋白的免疫荧光染色确定，玻璃上的细胞显示出更广泛的局灶性接触。流动下的细胞粘附减少硅烷通过抑制蛋白质合成与放线菌酮，这表明细胞附着到硅烷促进细胞合成Fn。结果表明，Fn细胞结合结构域的构象变化影响Fn对其细胞表面受体的亲和力。(C)John Wiley & Sons，Inc.
The effect of surface hydrophobicity upon the conformation of the cell binding domain of fibronectin (Fn) and the influence of Fn conformation on bovine aortic endothelial cell (BAEC) adhesion were examined. The free sulfhydryl group of Fn located near the cell binding domain was selectively labeled with acrylodan, a polarity sensitive fluor. Fluorescence emission was monitored in solution and upon adsorption to hydrophilic glass and hydrophobic silanized glass. The acrylodan-labeled Fn emission maximum shifted to longer wavelengths upon adsorption and the shift was greater for acrylodan-labeled Fn adsorbed to hydrophilic glass than hydrophobic silane, suggesting that the acrylodan was in a more solvent accessible environment on glass than silane. BAEC, suspended in serum-free medium, attached for 15 or 120 min onto glass or silane surfaces containing preadsorbed Fn, after which cell spreading and the strength of adhesion in a parallel plate flow chamber were measured. Cell spreading was similar on both surfaces after 15 min attachment, but BAECs were more spread on glass than silane after 120 min. At low surface concentrations of Fn, BAECs were more adherent on glass than silane. At higher surfaces concentrations, adhesion was similar. After a 2-h incubation in serum-free medium, cells on glass showed more extensive development of focal contacts as determined by immunofluorescent staining for vinculin. Cell adhesion under flow was reduced on silane by inhibition of protein synthesis with cycloheximide, suggesting that cell attachment to silane was promoted by cellular synthesis of Fn. The results indicate that changes in the conformation of the Fn cell binding domain affect Fn affinity for its cell surface receptor. (C) 1993 John Wiley & Sons, Inc.