Quantitative dynamics and binding studies of the 20S proteasome by NMR

摘要: The machinery used by the cell to perform essential biological processes is made up of large molecular assemblies. One such complex, proteasome, central machine for removal damaged and misfolded proteins from cell. Here we show that 670-kilodalton 20S proteasome core particle it possible overcome weight limitations have traditionally hampered quantitative nuclear magnetic resonance (NMR) spectroscopy studies systems. This achieved using an isotope labelling scheme where isoleucine, leucine valine methyls are protonated in otherwise highly deuterated background concert with experiments preserve lifetimes resulting NMR signals. methodology has been applied reveal functionally important motions interactions recording spectra on complexes weights a megadalton. Our results establish can provide detailed insight into supra-molecular structures over order magnitude larger than those routinely studied generally applicable.