title: Analysing properties of proteasome inhibitors using kinetic and X-ray crystallographic studies.
abstract: The combination of X-ray crystallography and kinetic studies of proteasome:ligand complexes has proven to be an important tool in inhibitor analysis of this crucial protein degradation machinery. Here, we describe in detail the purification protocols, proteolytic activity assays, crystallisation methods, and structure determination for the yeast 20S proteasome (CP) in complex with its inhibitors. The fusion of these advanced techniques offers the opportunity to further optimise drugs which are already tested in different clinical phase studies, as well as to design new promising proteasome lead structures which might be suitable for their application in medicine, plant protection, and antibiotics.
authors.affiliation: Department of Biochemistry, Technische Universität München, Garching, Germany.
authors.forename: Nerea
authors.initials: N
authors.lastname: Gallastegui
authors.forename: Michael
authors.initials: M
authors.lastname: Groll
mesh_terms: D000970:Antineoplastic Agents / Q000032:analysis; D001897:Boronic Acids / Q000494:pharmacology; D000069286:Bortezomib; D018360:Crystallography, X-Ray / Q000379:methods*; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000032:analysis* / Q000737:chemistry*; D007783:Lactones / Q000494:pharmacology; D009101:Multiple Myeloma / Q000188:drug therapy; D009842:Oligopeptides / Q000494:pharmacology; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism; D061988:Proteasome Inhibitors; D011719:Pyrazines / Q000494:pharmacology; D011758:Pyrroles / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D013912:Threonine / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D000970:Antineoplastic Agents; D001897:Boronic Acids; D004791:Enzyme Inhibitors; D007783:Lactones; D009842:Oligopeptides; D061988:Proteasome Inhibitors; D011719:Pyrazines; D011758:Pyrroles; D013912:Threonine; D000069286:Bortezomib; C527966:delanzomib; C475865:marizomib; C524865:carfilzomib; D046988:Proteasome Endopeptidase Complex