CONSTRUCTING PROTEINS BY DOVETAILING UNPROTECTED SYNTHETIC PEPTIDES - BACKBONE-ENGINEERED HIV PROTEASE
CONSTRUCTING PROTEINS BY DOVETAILING UNPROTECTED SYNTHETIC PEPTIDES - BACKBONE-ENGINEERED HIV PROTEASE
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DOI：
10.1126/science.1566069
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发表时间：
1992-04-10
期刊：
影响因子：
56.9
通讯作者：
KENT, SBH
中科院分区：
文献类型：
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作者：
SCHNOLZER, M;KENT, SBH
骨架工程化的HIV-1蛋白酶是通过全化学合成方法制备的，该方法将连接两个肽的行为与产生类似物结构相结合。对应于HIV-1蛋白酶单体多肽链的两半的未保护的合成肽片段通过独特的相互反应性官能团（每个片段上一个）干净且高产率地连接。在6摩尔盐酸胍中进行连接，从而避免了受保护的肽段的有限溶解度，这是蛋白质化学合成的经典方法的主要问题。所得的完全活性的HIV-1蛋白酶类似物含有硫酯取代物，用于两个活性位点瓣中的每一个中Gly 51-Gly 52之间的天然肽键，已知该区域对氨基酸侧链的突变变化高度敏感。
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.