title: On terminal alkynes that can react with active-site cysteine nucleophiles in proteases.
abstract: Active-site directed probes are powerful in studies of enzymatic function. We report an active-site directed probe based on a warhead so far considered unreactive. By replacing the C-terminal carboxylate of ubiquitin (Ub) with an alkyne functionality, a selective reaction with the active-site cysteine residue of de-ubiquitinating enzymes was observed. The resulting product was shown to be a quaternary vinyl thioether, as determined by X-ray crystallography. Proteomic analysis of proteins bound to an immobilized Ub alkyne probe confirmed the selectivity toward de-ubiquitinating enzymes. The observed reactivity is not just restricted to propargylated Ub, as highlighted by the selective reaction between caspase-1 (interleukin converting enzyme) and a propargylated peptide derived from IL-1β, a caspase-1 substrate.
authors: Division of Cell Biology, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
authors: Reggy
authors: R
authors: Ekkebus
authors: Sander I
authors: SI
authors: van Kasteren
authors: Yogesh
authors: Y
authors: Kulathu
authors: Arjen
authors: A
authors: Scholten
authors: Ilana
authors: I
authors: Berlin
authors: Paul P
authors: PP
authors: Geurink
authors: Annemieke
authors: A
authors: de Jong
authors: Soenita
authors: S
authors: Goerdayal
authors: Jacques
authors: J
authors: Neefjes
authors: Albert J R
authors: AJ
authors: Heck
authors: David
authors: D
authors: Komander
authors: Huib
authors: H
authors: Ovaa
mesh_terms: D000480:Alkynes / Q000737:chemistry*; D020134:Catalytic Domain; D003545:Cysteine / Q000737:chemistry*; D010447:Peptide Hydrolases / Q000378:metabolism*
chemical_list: D000480:Alkynes; D010447:Peptide Hydrolases; D003545:Cysteine
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