Connective tissue growth factor: a cysteine-rich mitogen secreted by human vascular endothelial cells is related to the SRC-induced immediate early gene product CEF-10.
Connective tissue growth factor: a cysteine-rich mitogen secreted by human vascular endothelial cells is related to the SRC-induced immediate early gene product CEF-10.
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DOI：
10.1083/jcb.114.6.1285
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发表时间：
1991-09
期刊：
影响因子：
--
通讯作者：
Grotendorst GR
中科院分区：
文献类型：
--
作者：
Bradham DM;Igarashi A;Potter RL;Grotendorst GR
此前已有报道称，人脐静脉内皮细胞表达PDGF的A链和B链基因，并向培养基中分泌PDGF相关因子。抗人PDGF IgG亲和层析法从HUVE细胞条件培养基中纯化PDGF相关活性。用抗PDGF IgG和PDGF A链或B链肽特异性抗体结合趋化和有丝分裂实验对亲和纯化的蛋白进行免疫印迹分析，发现HUVE细胞分泌的主要PDGF免疫相关分子是一个约36-38 kD的单体，纯化的生物活性分子中只有不到10%是PDGF A链或B链肽。利用抗PDGF抗体在表达载体lambda gtl 1中筛选HUVE细胞cDNA文库，克隆并测序了一个具有开放阅读框的cDNA，该cDNA编码一个38-kD富含半胱氨酸的分泌蛋白，我们发现该蛋白是人血管内皮细胞分泌的主要PDGF相关丝裂原。该蛋白与v- src诱导的鸡胚成纤维细胞CEF-10 mRNA的翻译产物总体同源性为45%。我们将这种新的有丝分裂原命名为结缔组织生长因子。
Human umbilical vein endothelial (HUVE) cells have been previously reported to express the genes for the A and B chains of PDGF and to secrete PDGF-related factors into culture media. Antihuman PDGF IgG affinity chromatography was used to purify PDGF-related activity from HUVE cell-conditioned media. Immunoblot analysis of the affinity- purified proteins with anti-PDGF IgG and antibodies specific for the A or B chain peptides of PDGF combined with chemotactic and mitogenic assays revealed that the major PDGF immunorelated molecule secreted by HUVE cells is a monomer of approximately 36-38 kD and that less than 10% of the purified biologically active molecules are PDGF A or B chain peptides. Screening of an HUVE cell cDNA library in the expression vector lambda gtl 1 with the anti-PDGF antibody resulted in the cloning and sequencing of a cDNA with an open reading frame encoding a 38-kD cysteine-rich secreted protein which we show to be the major PDGF- related mitogen secreted by human vascular endothelial cells. The protein has a 45% overall homology to the translation product of the v- src-induced CEF-10 mRNA from chick embryo fibroblasts. We have termed this new mitogen connective tissue growth factor.