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A Billion Ubiquitin Variants to Probe and Modulate the UPS

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1 A Billion Ubiquitin Variants to Probe and Modulate the UPS
Larissa A. Canadeo, Jon M. Huibregtse  Molecular Cell  Volume 62, Issue 1, Pages 2-4 (April 2016) DOI: /j.molcel Copyright © 2016 Elsevier Inc. Terms and Conditions

2 Figure 1 Strategy for Isolation and Characterization of HECT E3 UbVs
A phage display library expressing ubiquitin variants (UbVs) altered at one or more residues on the surface of ubiquitin was used to screen 20 different HECT E3s for tight-binding UbVs. These were assayed in vitro for their effects on E3 auto-ubiquitylation: some inhibited, some modulated chain length, and some activated ubiquitylation. X-ray structures of two inhibitory and four activating UbVs were determined, which was illuminating with respect to mechanisms of HECT E3 regulation. When expressed and assayed in a cell migration assay, the biological effects of a subset of UbVs were consistent with the in vitro effects on their cognate HECT E3s. Molecular Cell  , 2-4DOI: ( /j.molcel ) Copyright © 2016 Elsevier Inc. Terms and Conditions

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