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Science 10 April 1992:
Vol. 256. no. 5054, pp. 221 - 225
DOI: 10.1126/science.1566069
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Articles

Science, Vol 256, Issue 5054, 221-225
Copyright © 1992 by American Association for the Advancement of Science

articles

Constructing proteins by dovetailing unprotected synthetic peptides: backbone-engineered HIV protease

M Schnolzer and SB Kent

Department of Cell Biology, Scripps Research Institute, La Jolla, CA 92037.

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.


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