Semisynthetic protein-based site-directed probes for identification and inhibition of active sites, and methods therefor
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United States of America Patent
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Issued Date -
Dec 15, 2005
app pub date -
Apr 25, 2003
filing date -
Apr 25, 2002
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Abstract
A C-terminally modified Ubiquitin (Ub) derivative) ubiquitin vinyl sulfone (UbVS), which is specific for deubiquitinating enzymes (DUBs), was synthesized as an active site directed probe that irreversibly modifies a subset of Ub C-terminal hydrolases (UCHs) and Ub specific processing proteases (UBPs), is provided. [.sup.125I]-UbVS modifies 6 of the 17 known and putative yeast deubiquitinating enzymes, namely Yuh1p, Ubp1p, Ubp2p, Ubp6p, Ubp12p and Ubp15p. In mammalian cells, a greater number of polypeptides is labeled, most of which are DUBs. An additional DUB that associates with the mammalian 26S proteasome, novel protein USP14, a mammalian homolog of yeast Ubp6p that is bound to the proteasome, is provided.
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