title: Small, N-terminal tags activate Parkin E3 ubiquitin ligase activity by disrupting its autoinhibited conformation.
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.
authors.affiliation: Protein Structure and Function laboratory, The London Research Institute of Cancer Research UK, Lincoln's Inn Fields Laboratories, London, United Kingdom.
authors.forename: Lynn
authors.initials: L
authors.lastname: Burchell
authors.forename: Viduth K
authors.initials: VK
authors.lastname: Chaugule
authors.forename: Helen
authors.initials: H
authors.lastname: Walden
mesh_terms: D015853:Cysteine Proteinase Inhibitors / Q000737:chemistry; D000939:Epitopes / Q000737:chemistry; D057809:HEK293 Cells; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000737:chemistry; D006801:Humans; D007976:Leupeptins / Q000737:chemistry; D009842:Oligopeptides; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation; D055550:Protein Stability; D020598:Proto-Oncogene Proteins c-myb / Q000737:chemistry; D044767:Ubiquitin-Protein Ligases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: D015853:Cysteine Proteinase Inhibitors; D000939:Epitopes; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007976:Leupeptins; D009842:Oligopeptides; D010455:Peptides; D020598:Proto-Oncogene Proteins c-myb; C085706:FLAG peptide; D044767:Ubiquitin-Protein Ligases; C111567:parkin protein; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde