Tsang M L, Zhou L, Zheng B L, Wenker J, Fransen G, Humphrey J, Smith J M, O'Connor-McCourt M, Lucas R, Weatherbee J A
R&D Systems, Minneapolis, MN 55413, USA.
Cytokine. 1995 Jul;7(5):389-97. doi: 10.1006/cyto.1995.0054.
Recombinant human transforming growth factor soluble receptor Type II (rhTGF-beta sRII) corresponding to the 159 amino acid extracellular domain of hTGF-beta RII has been expressed in insect cells using the baculovirus expression system or in a mouse myeloma cell line. N-terminal sequence analysis of the purified protein revealed the removal of the 23 amino acid signal peptide. In SDS-PAGE, the rhTGF-beta sRII resolves into multiple bands due to N-linked glycosylation. Recombinant hTGF-beta sRII is a TGF-beta antagonist and will inhibit the biological activities of TGF-beta 1, TGF-beta 3, and TGF-beta 5 on TGF-beta-responsive cell lines, such as murine HT-2 or human TF-1 with an ED50 of approximately 0.3 micrograms/mL. However, hTGF-beta sRII does not inhibit TGF-beta 2 bioactivities in these cell lines, suggesting that hTGF-beta RII has low affinity for TGF-beta 2. Polyclonal antibodies to hTGF-beta sRII have been produced in goats and purified on Protein-G affinity columns. This antibody can inhibit TGF-beta 1,2,3,5-dependent bioactivities on human cell lines such as TF-1. Additionally, this antibody has species cross-reactivity and will also inhibit TGF-beta-dependent bioactivities on murine cells.(ABSTRACT TRUNCATED AT 250 WORDS)
对应于人转化生长因子β受体II（hTGF-βRII）159个氨基酸胞外域的重组人转化生长因子可溶性受体II型（rhTGF-β sRII）已利用杆状病毒表达系统在昆虫细胞中或在小鼠骨髓瘤细胞系中表达。对纯化蛋白进行N端序列分析显示去除了23个氨基酸的信号肽。在SDS-PAGE中，由于N-糖基化，rhTGF-β sRII可分离为多条带。重组hTGF-β sRII是一种TGF-β拮抗剂，将抑制TGF-β 1、TGF-β 3和TGF-β 5对TGF-β反应性细胞系（如鼠源HT-2或人源TF-1）的生物学活性，ED50约为0.3微克/毫升。然而，hTGF-β sRII在这些细胞系中不抑制TGF-β 2的生物学活性，这表明hTGF-β RII对TGF-β 2的亲和力较低。已在山羊中制备了针对hTGF-β sRII的多克隆抗体，并在Protein-G亲和柱上进行了纯化。该抗体可抑制TGF-β 1、2、3、5对人细胞系（如TF-1）的依赖性生物学活性。此外，该抗体具有种间交叉反应性，也将抑制TGF-β对鼠细胞的依赖性生物学活性。（摘要截短于250字）
