Protocol

Ubiquitin Family Modifiers and the Proteasome

Volume 832 of the series Methods in Molecular Biology pp 373-390

Date:

Analysing Properties of Proteasome Inhibitors Using Kinetic and X-Ray Crystallographic Studies

  • Nerea GallasteguiAffiliated withDepartment of Biochemistry, Technische Universität München
  • , Michael GrollAffiliated withDepartment of Biochemistry, Technische Universität München Email author 

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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.

Key words

Proteasome Crystallography Kinetic studies Inhibitors Drug design Cancer