Schnölzer M, Kent S B
Department of Cell Biology, Scripps Research Institute, La Jolla, CA 92037.
Science. 1992 Apr 10;256(5054):221-5. doi: 10.1126/science.1566069.
Backbone-engineered HIV-1 protease was prepared by a total chemical synthesis approach that combines the act of joining two peptides with the generation of an analog structure. Unprotected synthetic peptide segments corresponding to the two halves of the HIV-1 protease monomer polypeptide chain were joined cleanly and in high yield through unique mutually reactive functional groups, one on each segment. Ligation was performed in 6 molar guanidine hydrochloride, thus circumventing limited solubility of protected peptide segments, the principal problem of the classical approach to the chemical synthesis of proteins. The resulting fully active HIV-1 protease analog contained a thioester replacement for the natural peptide bond between Gly51-Gly52 in each of the two active site flaps, a region known to be highly sensitive to mutational changes of amino acid side chains.
通过一种全化学合成方法制备了主链工程化的HIV-1蛋白酶，该方法将两个肽段的连接行为与类似物结构的生成相结合。与HIV-1蛋白酶单体多肽链两半相对应的未保护合成肽段通过独特的相互反应性功能基团干净且高产率地连接在一起，每个肽段上各有一个这样的基团。连接反应在6摩尔盐酸胍中进行，从而避免了受保护肽段溶解度有限的问题，而这是经典蛋白质化学合成方法的主要问题。所得的具有完全活性的HIV-1蛋白酶类似物在两个活性位点侧翼中每个的Gly51 - Gly52之间的天然肽键处含有硫酯替代物，该区域已知对氨基酸侧链的突变变化高度敏感。
