Recombinant ATPases of the yeast 26S proteasome activate protein degradation by the 20S proteasome.
The 26S proteasome contains a proteolytic core, 20S proteasome, and its regulatory particle, 19S complex. That regulatory particle contains six ATPases that are involved in unfolding and translocation of substrates to the 20S proteasome's catalytic chamber. We expressed ATPase-encoding genes of the regulatory particle of Saccharomyces cerevisiae and found that some recombinant ATPases can self-assemble into a high-molecular-weight protein complex in Escherichia coli. Purification of the Rpt1Rpt2 hetero-complex and the Rpt4 homo-complex for functional characterization demonstrated their contribution to energy-dependent protein degradation. Our finding, production of a functional subunit of the 19S regulatory particle in bacteria, is a simpler and technically advanced system to functionally characterize individual subunits.
Takeuchi
Junko
J
Proteolysis and Protein Turnover Research Group, Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Toyohira-ku, Sapporo 062-8517, Japan.
Tamura
Tomohiro
T
D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000502:physiology; D000255:Adenosine Triphosphate / Q000737:chemistry; D002384:Catalysis; D003546:Cysteine Endopeptidases / Q000737:chemistry*; D004268:DNA-Binding Proteins / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000737:chemistry* / Q000502:physiology; D004926:Escherichia coli / Q000378:metabolism; D006868:Hydrolysis; D009097:Multienzyme Complexes / Q000737:chemistry*; D010447:Peptide Hydrolases / Q000737:chemistry*; D010957:Plasmids / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex; D017510:Protein Folding; D021381:Protein Transport; D011994:Recombinant Proteins / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000502:physiology; D029702:Schizosaccharomyces pombe Proteins / Q000502:physiology; D013997:Time Factors
D004268:DNA-Binding Proteins; D009097:Multienzyme Complexes; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D029702:Schizosaccharomyces pombe Proteins; D000255:Adenosine Triphosphate; D010450:Endopeptidases; D010447:Peptide Hydrolases; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease; C084436:rpt2 protein, S pombe; C119011:26S proteasome non-ATPase regulatory subunit 13; D000251:Adenosine Triphosphatases; C103015:RPT4 protein, S cerevisiae; C084493:RPT1 protein, S cerevisiae