Thrombin peptide, TP508, induces differential gene expression in fibroblasts through a nonproteolytic activation pathway.
Thrombin peptide, TP508, induces differential gene expression in fibroblasts through a nonproteolytic activation pathway.
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凝血酶肽 TP508 通过非蛋白水解激活途径诱导成纤维细胞差异基因表达。
DOI：
10.1006/excr.1998.4372
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Carney,DH
中科院分区：
文献类型：
--
作者：
Sower,LE;Payne,DA;Meyers,R;Carney,DH
先前的研究表明，合成肽代表凝血酶结构域，负责与成纤维细胞的高亲和力结合，通过非蛋白分解机制刺激趋化和细胞增殖信号。其中一种多肽，TP508，最近被证明对中性粒细胞具有趋化作用，促进伤口中胶原的积累，促进伤口的血管重建，并加速正常动物和愈合受损动物的切开创面和开放创面的愈合。为了确定TP508是否激活蛋白水解性凝血酶受体(PAR1)或PAR1激活的信号，我们用TP508和PAR1激活肽SFLLRNP处理人成纤维细胞，并用差异显示逆转录聚合酶链式反应分析这些肽对基因表达的影响。TP508可诱导SFLLRNP不能诱导的具有短酪氨酸激酶样结构域的大量特异信息片段的表达。通过对TP508诱导片段进行马拉松延伸获得的全长克隆测序表明，在诱导的转录本中，有一个序列与人Annexin V有88%的同源性。Northern分析证实，TP508而不是SFLLRNP能够诱导人成纤维细胞表达Annexin V。这些结果表明，TP508激活了不同于通过PAR1激活的细胞反应，并支持了TP508通过单独的非蛋白水解性激活的凝血酶受体发挥作用的假设，该受体可能负责高亲和力凝血酶结合和凝血酶刺激细胞增殖所需的非蛋白水解性信号。
Prior studies have shown that synthetic peptides representing the domain of thrombin responsible for high-affinity binding to fibroblasts stimulate chemotactic and cell proliferative signals through a nonproteolytic mechanism. One of these peptides, TP508, has recently been shown to be chemotactic for neutrophils, to enhance collagen accumulation in wounds, to enhance revascularization of wounds, and to accelerate the healing of incisional and open wounds in normal animals and in animals with impaired healing. To determine whether TP508 activates the proteolytically activated receptor for thrombin (PAR1), or the signals that are activated by PAR1, we treated human fibroblasts with TP508 and the PAR1-activating peptide, SFLLRNP, and analyzed the effects of these peptides on gene expression using differential display reverse transcriptase polymerase chain reaction. TP508 induces expression of a number of specific message fragments with short tyrosine kinase-like domains that are not induced by SFLLRNP. Sequencing full-length clones prepared by Marathon extension of TP508-induced fragments revealed that among the induced transcripts, there was a sequence with 88% homology to human annexin V. Northern analysis with authentic annexin V cDNA confirms that TP508, but not SFLLRNP, induces expression of annexin V in human fibroblasts. These results demonstrate that TP508 activates a cellular response separate from that activated through PAR1 and supports the hypothesis that TP508 acts through a separate nonproteolytically activated thrombin receptor that may be responsible for high-affinity thrombin binding and for nonproteolytic signals that are required for thrombin stimulation of cell proliferation.