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Preparation of peptide thioesters from naturally occurring sequences using reaction sequence consisting of regioselective S‐cyanylation and hydrazinolysis
Author(s) -
Miyajima Rin,
Tsuda Yusuke,
Inokuma Tsubasa,
Shigenaga Akira,
Imanishi Miki,
Futaki Shiroh,
Otaka Akira
Publication year - 2016
Publication title -
peptide science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.22757
Subject(s) - thioester , native chemical ligation , chemistry , cysteine , peptide , regioselectivity , chemical ligation , combinatorial chemistry , hydrazide , chemical synthesis , chemical space , residue (chemistry) , chemical modification , biochemistry , organic chemistry , drug discovery , enzyme , in vitro , catalysis
ABSTRACT The vital roles of peptide/protein thioesters in protein chemistry, including chemical or semi‐synthesis of proteins, have encouraged studies on the development of methods for the preparation of such chemical units. Biochemical protocols using intein or sortase have proved to be useful in protein chemistry as methods suitable for naturally occurring sequences, including recombinant proteins. Although chemical protocols are potential options for thioester preparation, only a few are applicable to naturally occurring sequences, because standard chemical protocols require an artificial chemical device for producing thioesters. In this context, the chemical preparation of thioesters based on a reaction sequence consisting of regioselective S‐cyanylation and hydrazinolysis was investigated. Regioselective S‐cyanylation, which is required for cysteine‐containing thioesters, was achieved with the aid of a zinc‐complex formation of a CCHH‐type zinc‐finger sequence. Free cysteine residues that are not involved in complex formation were selectively protected with a 6‐nitroveratryl group followed by S‐cyanylation of the zinc‐binding cysteine. Hydrazinolysis of the resulting S‐cyanopeptide and subsequent photo‐removal of the 6‐nitroveratryl group yielded the desired peptide hydrazide, which was then converted to the corresponding thioester. The generated thioester was successfully used in N–to–C‐directed one‐pot/sequential native chemical ligation using an N‐sulfanylethylanilide peptide to give a 64‐residue peptide toxin. © 2015 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 106: 531–546, 2016.