Purification of Proteasome Subcomplexes

Endogenous holoenzyme, core particle ^{41 (link)}, and lid subcomplex ^{42 (link)} were purified from S. cerevisiae essentially as described. The base subcomplex was purified according to protocols for the holoenzyme preparation, but with minor modifications as described in the Full Methods. Details of yeast strain construction are provided in Table S1.
Yeast lid was recombinantly expressed from three plasmids in E. coli BL21-star (DE3), and purified on anti-FLAG M2 resin and by size-exclusion chromatography (see Full Methods).

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Lander G.C., Estrin E., Matyskiela M.E., Bashore C., Nogales E, & Martin A. (2012). Complete subunit architecture of the proteasome regulatory particle. Nature, 482(7384), 186-191.

Publication 2011

E coli Holoenzyme Plasmids Resin Size exclusion chromatography Strain Yeast

Corresponding Organization :

Other organizations : Lawrence Berkeley National Laboratory, University of California, Berkeley, Howard Hughes Medical Institute

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Variable analysis

independent variables

Endogenous holoenzyme, core particle
Lid subcomplex
Recombinant expression of yeast lid in E. coli BL21-star (DE3)

dependent variables

Purification of endogenous holoenzyme, core particle, and lid subcomplex from S. cerevisiae
Purification of recombinant yeast lid from E. coli

control variables

Yeast strain construction details provided in Table S1

positive controls

Holoenzyme, core particle, and lid subcomplex purified from S. cerevisiae as described in previous studies

negative controls

Not explicitly mentioned

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