small, n-terminal tags activate parkin e3 ubiquitin ligase activity by disrupting its autoinhibited conformation小,n端标签激活帕金e3泛素连接酶活性通过破坏其autoinhibited构象.pdfVIP

Small, N-Terminal Tags Activate Parkin E3 Ubiquitin Ligase Activity by Disrupting Its Autoinhibited Conformation ¤ Lynn Burchell, Viduth K. Chaugule , Helen Walden* Protein Structure and Function laboratory, The London Research Institute of Cancer Research UK, Lincoln’s Inn Fields Laboratories, London, United Kingdom Abstract Parkin is an E3 ubiquitin ligase, mutations in which cause Autosomal Recessive Parkinson’s Disease. Many studies aimed at understanding Parkin function, regulation and dysfunction are performed using N-terminal epitope tags. We report here that the use of small tags such as FLAG, cMyc and HA, influence the physical stability and activity of Parkin in and out of cells, perturbing the autoinhibited native state of Parkin, resulting in an active-for-autoubiquitination species. Citation: Burchell L, Chaugule VK, Walden H (2012) Small, N-Terminal Tags Activate Parkin E3 Ubiquitin Ligase Activity by Disrupting Its Autoinhibited Conformation. PLoS ONE 7(4): e34748. doi:10.1371/journal.pone.0034748 Editor: Alexander J. Whitworth, University of Sheffield - MRC Centre for Developmental and Biomedical Genetics, United Kingdom Received January 13, 2012; Accepted March 8, 2012; Published April 4, 2012 Copyright: 2012 Burchell et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This work was funded by Cancer Research UK. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. * E-mail: Helen.walden@.uk ¤ Current address: Department of Epige