Chang H H, Hu S T, Huang T F, Chen S H, Lee Y H, Lo S J
Graduate Institute of Microbiology and Immunology, National Yang-Ming Medical College, Taipei, Taiwan.
Biochem Biophys Res Commun. 1993 Jan 15;190(1):242-9. doi: 10.1006/bbrc.1993.1037.
Rhodostomin (Rho) from snake venom, a potent inhibitor of platelet aggregation, contains 68 amino acids having an RGD sequence and 12 cysteine residues. A chemically synthesized Rho gene was cloned and expressed in Escherichia coli. The expression of Rho gene fused with the glutathione S-transferase (GST) gene was about 10-30% of total cell proteins. The Rho-fusion protein could be recognized by antibodies raised against either a native Rho peptide or a synthetic peptide. The purified GST-Rho coated on culture plates facilitated the attachment of human hepatoma cells, which was inhibitable by co-incubation with a synthetic hexapeptide GRGDSP but not with a related peptide of GRGESP, suggesting that the E. coli-expressed Rho-fusion protein was properly folded and biologically functional.
来自蛇毒的罗多斯汀（Rho）是一种有效的血小板聚集抑制剂，含有68个氨基酸，具有RGD序列和12个半胱氨酸残基。一个化学合成的Rho基因被克隆并在大肠杆菌中表达。与谷胱甘肽S-转移酶（GST）基因融合的Rho基因的表达量约占总细胞蛋白的10%-30%。Rho融合蛋白可被针对天然Rho肽或合成肽产生的抗体识别。包被在培养板上的纯化的GST-Rho促进了人肝癌细胞的附着，与合成六肽GRGDSP共同孵育可抑制这种附着，但与相关肽GRGESP共同孵育则不能，这表明大肠杆菌表达的Rho融合蛋白正确折叠且具有生物学功能。
