Monson J M
Coll Relat Res. 1983;3(1):1-12. doi: 10.1016/s0174-173x(83)80044-2.
Proteolytic digestion was used to probe the conformation of the prepro alpha chains synthesized by a mRNA-dependent reticulocyte lysate from chicken calvarial RNA. Pepsin-resistant alpha 1- and alpha 2-like chains were recovered even from translation reactions that were not preincubated below the reported Tm of the unhydroxylated triple helix. The pepsin-resistant structures were stable to thermal denaturation at 45 degrees C and a fraction remained resistant to peptic digestion at 30 degrees C. Interchain disulfide bonds did not appear to be required for the formation or thermal stability of these structures. Pepsin resistance is normally interpreted as evidence for a triple-helical conformation. Therefore, these results suggest that the in vitro synthesized prepro alpha chains contain the requisite information to associate in register for correct helix folding. The unusual thermal stability of these structures is not understood, but this may indicate assembly into higher orders of structure.
采用蛋白水解消化法来探究由鸡颅骨RNA的mRNA依赖性网织红细胞裂解物合成的前原α链的构象。即使在未低于报道的未羟基化三螺旋的熔解温度（Tm）进行预孵育的翻译反应中，也能回收抗胃蛋白酶的α1和α2样链。这些抗胃蛋白酶的结构在45℃下对热变性稳定，并且一部分在30℃下仍能抵抗胃蛋白酶消化。链间二硫键似乎不是这些结构形成或热稳定性所必需的。抗胃蛋白酶性通常被解释为三螺旋构象的证据。因此，这些结果表明体外合成的前原α链包含正确螺旋折叠所需的对齐结合信息。这些结构异常的热稳定性尚不清楚，但这可能表明它们组装成了更高阶的结构。
