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One‐Step Chemical Synthesis of Native Met1‐Linked Poly‐Ubiquitin Chains
Author(s) -
van der Heden van Noort Gerbrand J.,
Talavera Ormeño Cami,
van Dalen Duco,
Ovaa Huib
Publication year - 2019
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201800520
Subject(s) - ubiquitin , native chemical ligation , regioselectivity , chemistry , chemical ligation , folding (dsp implementation) , lysine , deubiquitinating enzyme , enzyme , ubiquitin ligase , biochemistry , combinatorial chemistry , amino acid , cysteine , catalysis , electrical engineering , gene , engineering
The enzyme‐mediated construction of poly‐ubiquitin (Ub) chains on target proteins leads to a variety of cellular responses and is involved in processes ranging from protein degradation to cell cycle control and immune responses. This complex post‐translational modification system is under intense investigation, but generation of specific Ub chains and tools made thereof is not always trivial. We discovered that native methionine‐1‐linked polymeric ubiquitin chains can be constructed in a single chemical reaction. We validate correct folding and regioselective attachment of such chains using linkage specific proteases and further demonstrate that these poly‐Ub chains can be converted into thioesters by the activating E1‐enzyme. Subsequent ligation reactions using these in situ prepared thioesters leads to poly‐ubiquitinated peptides.