Synthesis of biologically active transforming growth factor alpha.
Synthesis of biologically active transforming growth factor alpha.
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生物活性转化生长因子α的合成。
DOI：
10.1111/j.1399-3011.1987.tb02269.x
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
Tam,JP
中科院分区：
文献类型：
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作者：
Tam,JP
通过改进的分步固相法合成了由猫肉瘤病毒转化的大鼠胚胎成纤维细胞在培养物中分泌的50个氨基酸残基的转化生长因子α（TGFα），总产率为31%。采用基于SN 2机制的脱保护策略，使用低浓度的HF或CF 3SO 3 H-CF 3CO 2 H的二甲硫醚溶液来去除大部分苄基衍生的保护基。通过使用高浓度HF的SN 2条件除去Cys和Arg的更耐酸的保护基团。合成的TGFα在三个步骤中纯化至均一。合成和天然TGFα在HPLC和不同试验中无法区分，包括正常大鼠肾成纤维细胞在软琼脂中的锚定非依赖性生长、结合和刺激表皮生长因子（EGF）受体蛋白激酶的试验。此外，在这些测定中，合成的TGFα与EGF相比显示出相似的生物活性。因此，TGFα的化学合成提供了令人信服的证据，表明TGFα在功能上与EGF相关，并且是细胞转化所需的活性成分之一。
A 50‐amino acid residue transforming growth factor, type alpha (TGFα), secreted in culture by feline‐sarcoma‐virus‐transformed rat embryo fibroblasts, was synthesized by an improved stepwise solid‐phase method with an overall yield of 31%. A deprotection strategy based on the SN2 mechanism using either a low concentration of HF or CF3SO3H‐CF3CO2H in dimethylsulfide was employed to remove most of the benzyl‐derived protecting groups. The more acid resistant protecting groups of Cys and Arg were removed by the SN2 condition using a high concentration of HF. Synthetic TGFα was purified to homogeneity in three steps. Synthetic and natural TGFα were indistinguishable from each other in HPLC and in different assays, including the assay for anchorage‐independent growth of normal rat kidney fibroblasts in soft agar, binding, and stimulating to epidermal growth factor (EGF)‐receptor protein kinase. Furthermore, synthetic TGFα showed similar biological activities when compared with EGF in these assays. Thus, the chemical synthesis of TGFα provided convincing evidence that TGFα is functionally related to EGF and is one of the active principles required for cellular transformation.