VASCULAR ENDOTHELIAL-CELL GROWTH-FACTOR (VEGF) PRODUCED BY A-431 HUMAN EPIDERMOID CARCINOMA-CELLS AND IDENTIFICATION OF VEGF MEMBRANE-BINDING SITES
VASCULAR ENDOTHELIAL-CELL GROWTH-FACTOR (VEGF) PRODUCED BY A-431 HUMAN EPIDERMOID CARCINOMA-CELLS AND IDENTIFICATION OF VEGF MEMBRANE-BINDING SITES
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DOI：
10.1073/pnas.88.13.5819
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发表时间：
1991-07-01
影响因子：
11.1
通讯作者：
SATO, JD
中科院分区：
文献类型：
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作者：
MYOKEN, Y;KAYADA, Y;SATO, JD
内皮细胞有丝分裂原的一个独特的家族，是同源的血小板衍生的生长因子最近被确定。与其他已知的内皮细胞有丝分裂原不同，这些血管内皮细胞生长因子（vegf）是分泌的，并且似乎对内皮细胞有特异性作用。我们从A-431人表皮样癌细胞培养的无蛋白培养基中纯化了2083倍的VEGF，达到明显的均匀性。该a -431来源的VEGF是一种同源二聚体，由22 kda亚基组成，其n端序列与人HL-60白血病和U-937组织细胞淋巴瘤细胞产生的VEGF相似。A-431 VEGF用于鉴定VEGF在人脐静脉内皮细胞（HUVEC）上的特异性和饱和结合位点。通过亲和交联，在HUVEC上检测到230、170和125 kDa的vegf结合位点复合物。VEGF特异性诱导了一个190 kda多肽的酪氨酸磷酸化，该多肽的质量与亲和交联检测到的最大结合位点相似。
A distinct family of endothelial cell mitogens that are homologous to platelet-derived growth factor has recently been identified. Unlike other known endothelial cell mitogens, these vascular endothelial cell growth factors (VEGFs) are secreted and appear to act specifically on endothelial cells. We have purified VEGF 2083-fold to apparent homogeneity from protein-free culture medium conditioned by A-431 human epidermoid carcinoma cells. This A-431-derived VEGF was characterized as a homodimer composed of 22-kDa subunits with an N-terminal sequence that was similar to VEGFs produced by human HL-60 leukemic and U-937 histiocytic lymphoma cells. A-431 VEGF was used to identify specific and saturable binding sites for VEGF on human umbilical vein endothelial cells (HUVEC). By affinity cross-linking, VEGF-binding site complexes of 230, 170, and 125 kDa were detected on HUVEC. VEGF specifically induced the tyrosine phosphorylation of a 190-kDa polypeptide, which was similar in mass to the largest binding site detected by affinity cross-linking.