Chemical synthesis of TASP arrays and their application in protein design

Molecular Diversity
Wolfgang Haehnel

Abstract

A combinatorial synthesis of de novo proteins is described. The concept of template-assembled synthetic proteins (TASP) has been adapted to an orthogonal assembly of small libraries of purified peptide building blocks. It is combined with the spot synthesis of peptides which is exploited to array cyclic decapeptide templates on cellulose membranes. A cleavable linker on the cellulose allows control of the synthesis. The hydrophilic proteins are constructed by successive cleavage of orthogonal protecting groups on the template, followed by coupling of amphipathic helices in a predefined orientation and finally by incorporation of a cofactor. Libraries of peptides with variation of the amino acids expected to be close to the cofactor were coupled to the cellulose-bound template in all combinations, yielding up to 500 variants of a protein. Cofactors have been inserted either at non-covalent binding sites as heme and Cu2+ or by covalent modification of amino acids as Ru-bipyridine or flavin. The proteins were screened by recording their UV-vis spectra directly on the solid support. The properties screened include the redox potential of heme proteins, charge transfer bands indicating the ligation of Cu-centers, enzymatic activity, ...Continue Reading

Citations

Nov 13, 2008·Proteins·Zhong Zheng, M R Gunner
May 18, 2016·Organic & Biomolecular Chemistry·Frédérique DeissRatmir Derda
Nov 22, 2007·Journal of Peptide Science : an Official Publication of the European Peptide Society·Didier BoturynPascal Dumy
May 21, 2008·Proteins·Christian FufezanM R Gunner
Mar 17, 2005·Chemistry : a European Journal·Pradip Chakraborty, Ulf Diederichsen
Aug 8, 2006·Chembiochem : a European Journal of Chemical Biology·Yashveer SinghPascal Dumy
May 14, 2009·Chembiochem : a European Journal of Chemical Biology·Rudolf Volkmer
Jul 5, 2006·Chemistry : a European Journal·Wen-Wu LiWolfgang Haehnel
Dec 7, 2007·Biotechnology & Genetic Engineering Reviews·Kai HilperRobert E W Hancock

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