TY - JOUR
T1 - Expressed Protein Selenoester Ligation
AU - Kulkarni, Sameer S.
AU - Watson, Emma E.
AU - Maxwell, Joshua W.C.
AU - Niederacher, Gerhard
AU - Johansen-Leete, Jason
AU - Huhmann, Susanne
AU - Mukherjee, Somnath
AU - Norman, Alexander R.
AU - Kriegesmann, Julia
AU - Becker, Christian F.W.
AU - Payne, Richard J.
N1 - Publisher Copyright:
© 2022 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.
PY - 2022/5/9
Y1 - 2022/5/9
N2 - Herein, we describe the development and application of a novel expressed protein selenoester ligation (EPSL) methodology for the one-pot semi-synthesis of modified proteins. EPSL harnesses the rapid kinetics of ligation reactions between modified synthetic selenopeptides and protein aryl selenoesters (generated from expressed intein fusion precursors) followed by in situ chemoselective deselenization to afford target proteins at concentrations that preclude the use of traditional ligation methods. The utility of the EPSL technology is showcased through the efficient semi-synthesis of ubiquitinated polypeptides, lipidated analogues of the membrane-associated GTPase YPT6, and site-specifically phosphorylated variants of the oligomeric chaperone protein Hsp27 at high dilution.
AB - Herein, we describe the development and application of a novel expressed protein selenoester ligation (EPSL) methodology for the one-pot semi-synthesis of modified proteins. EPSL harnesses the rapid kinetics of ligation reactions between modified synthetic selenopeptides and protein aryl selenoesters (generated from expressed intein fusion precursors) followed by in situ chemoselective deselenization to afford target proteins at concentrations that preclude the use of traditional ligation methods. The utility of the EPSL technology is showcased through the efficient semi-synthesis of ubiquitinated polypeptides, lipidated analogues of the membrane-associated GTPase YPT6, and site-specifically phosphorylated variants of the oligomeric chaperone protein Hsp27 at high dilution.
KW - Expressed Protein Selenoesters
KW - Peptides
KW - Protein Modifications
KW - Protein Semi-Synthesis
KW - Proteins
UR - http://www.scopus.com/inward/record.url?scp=85126051441&partnerID=8YFLogxK
U2 - 10.1002/anie.202200163
DO - 10.1002/anie.202200163
M3 - Article
C2 - 35194928
AN - SCOPUS:85126051441
SN - 1433-7851
VL - 61
JO - Angewandte Chemie - International Edition
JF - Angewandte Chemie - International Edition
IS - 20
M1 - e202200163
ER -