title: Controlled enzymatic synthesis of natural-linkage, defined-length polyubiquitin chains using lysines with removable protecting groups.
abstract: E2 enzymes catalyze the ATP-dependent polymerization of polyubiquitin chains which function as molecular signals in the regulation of numerous cellular processes. Here we present a method that uses genetically encoded unnatural amino acids to halt and re-start ubiquitin polymerization providing access to natural-linkage, precision-length ubiquitin chains that can be used for biochemical, structural, and dynamics studies.
authors.affiliation: Center for Biomolecular Structure and Organization, University of Maryland, College Park, Maryland 20742, USA.
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Castañeda
authors.forename: Jia
authors.initials: J
authors.lastname: Liu
authors.forename: Tanuja R
authors.initials: TR
authors.lastname: Kashyap
authors.forename: Rajesh K
authors.initials: RK
authors.lastname: Singh
authors.forename: David
authors.initials: D
authors.lastname: Fushman
authors.forename: T Ashton
authors.initials: TA
authors.lastname: Cropp
mesh_terms: D004926:Escherichia coli / Q000235:genetics; D008239:Lysine / Q000031:analogs & derivatives* / Q000235:genetics; D009154:Mutation; D058105:Polymerization; D025821:Polyubiquitin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D025801:Ubiquitin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D044763:Ubiquitin-Conjugating Enzymes / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D025801:Ubiquitin; D025821:Polyubiquitin; D044763:Ubiquitin-Conjugating Enzymes; D008239:Lysine