Abiko T, Sekino H
Kidney Research Laboratory, Sendai, Japan.
Biotechnol Ther. 1993;4(3-4):213-20.
The complete murine prothymosin alpha molecule (110 residues) except for the N-terminal methionine deduced from the cloned cDNA has been synthesized by a solid-phase method. Peptide synthesis was performed manually by the stepwise solid-phase method using the base-labile Fmoc group for protecting the alpha-amino group. The peptide was assembled on a p-alkoxybenzyl alcohol resin. After the last coupling step, the Fmoc group was removed with 50% piperidine in DMF. The peptide resin was treated with thioanisole-o-cresol in TFA, and then purified by gel filtration, ion-exchange column chromatography and high-performance liquid chromatography. A 2.9-mg sample of a highly purified peptide was finally obtained. The overall yield of the synthesis was less than 1%, based on the amino acid content of the starting Fmoc-Asp (OtBu)-resin. The synthetic peptide was found to have a restoring activity on low-E-rosette-forming lymphocytes after incubation of peripheral blood from uremic patients with the synthetic peptide. This peptide exhibited far stronger restoring effect than that of our synthetic thymosin alpha 1.
已通过固相法合成了完整的鼠源前胸腺素α分子（110个残基），该分子是根据克隆的cDNA推导而来，N端甲硫氨酸除外。肽合成通过逐步固相法手动进行，使用对碱不稳定的Fmoc基团保护α-氨基。肽在对烷氧基苄醇树脂上组装。在最后一个偶联步骤后，用50%哌啶的DMF溶液除去Fmoc基团。肽树脂用硫代苯甲醚-邻甲酚的TFA溶液处理，然后通过凝胶过滤、离子交换柱色谱和高效液相色谱进行纯化。最终获得了2.9mg高纯度肽样品。基于起始Fmoc-Asp（OtBu）-树脂的氨基酸含量，合成的总产率低于1%。将尿毒症患者的外周血与合成肽孵育后，发现合成肽对低E-玫瑰花结形成淋巴细胞具有恢复活性。该肽表现出比我们合成的胸腺素α1更强的恢复效果。
