Marquardt H, Hunkapiller M W, Hood L E, Twardzik D R, De Larco J E, Stephenson J R, Todaro G J
Proc Natl Acad Sci U S A. 1983 Aug;80(15):4684-8. doi: 10.1073/pnas.80.15.4684.
Transforming growth factors (TGFs) were purified from serum-free medium conditioned by retrovirus-transformed Fisher rat embryo fibroblasts, mouse 3T3 cells, and two human melanoma cell lines. The purification of each TGF was monitored in a radioreceptor assay based on receptor crossreactivity with mouse submaxillary gland epidermal growth factor (mEGF) and was achieved by gel permeation chromatography of the acid-soluble TGF-containing activity, followed by reverse-phase high-pressure liquid chromatography with sequential use of acetonitrile and 1-propanol in the presence of aqueous trifluoroacetic acid. The amino-terminal sequences of rat, mouse, and human TGFs were determined. Extensive sequence homology was found among TGF polypeptides from different species and cell types. Alignment of the amino acid sequences of rat TGF, mEGF, and human urogastrone (hEGF) reveals statistically significant sequence homology. The reported results suggest that TGFs that compete for binding to the cellular EGF receptor and EGF may have evolved from a common progenitor.
转化生长因子（TGFs）是从无血清培养基中纯化得到的，该培养基由逆转录病毒转化的Fisher大鼠胚胎成纤维细胞、小鼠3T3细胞和两个人类黑色素瘤细胞系培养而来。基于与小鼠颌下腺表皮生长因子（mEGF）的受体交叉反应性，通过放射受体测定法监测每种TGF的纯化过程。通过对含酸溶性TGF活性进行凝胶渗透色谱，然后在三氟乙酸水溶液存在下依次使用乙腈和1-丙醇进行反相高压液相色谱，实现了TGF的纯化。测定了大鼠、小鼠和人类TGF的氨基末端序列。发现来自不同物种和细胞类型的TGF多肽之间存在广泛的序列同源性。大鼠TGF、mEGF和人类尿抑胃素（hEGF）氨基酸序列的比对显示出具有统计学意义的序列同源性。报道的结果表明，竞争与细胞表皮生长因子受体结合的TGFs和表皮生长因子可能起源于共同的祖先。
