Simultaneous binding of PA28 and PA700 activators to 20 S proteasomes.
Two activators, named PA700 and PA28, are known to bind to 20 S proteasomes, forming two different complexes. The PA700-proteasome complex, also known as the 26 S proteasome, can degrade intact proteins, whereas complexes with PA28 can degrade only peptides. Monoclonal antibodies to 20 S proteasomes or the p45 ATPase subunit (Trip1, Sug1) of PA700 precipitated the same set of proteins from HeLa extracts, including six different ATPase subunits of PA700. This shows that p45 is not present in other protein complexes and suggests that all 26 S proteasome particles contain the same set of ATPase subunits. Interferons alpha and gamma had no effect on the composition of the 26 S proteasome, except for the replacement of subunits delta, MB1 and Z with Lmp2, Lmp7 and MECL1 respectively. Surprisingly, antibodies to PA28 precipitated p42, a component of PA700. Conversely, anti-p45 antibodies precipitated not only 26 S proteasomes but also PA28 alpha, beta and gamma, indicating that 20 S proteasomes can simultaneously bind both PA700 and PA28. PA28 alpha beta is known to be involved in antigen presentation. Conceivably, intact substrate proteins are recognized by PA700 and fed into proteasomes whose cleavage specificity is optimized for antigen presentation on MHC class I by PA28 and three interferon inducible proteasome subunits.
Hendil
K B
KB
August Krogh Institute, University of Copenhagen, Universitetsparken 13, DK-2100 Copenhagen O, Denmark. KBHendil@aki.ku.dk
Khan
S
S
Tanaka
K
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D000251:Adenosine Triphosphatases / Q000276:immunology / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000276:immunology; D002850:Chromatography, Gel; D003546:Cysteine Endopeptidases / Q000276:immunology / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D004789:Enzyme Activation; D006367:HeLa Cells; D006801:Humans; D016898:Interferon-alpha / Q000378:metabolism; D007371:Interferon-gamma / Q000378:metabolism; D009097:Multienzyme Complexes / Q000276:immunology / Q000378:metabolism*; D009124:Muscle Proteins; D010447:Peptide Hydrolases / Q000276:immunology / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D016898:Interferon-alpha; D009097:Multienzyme Complexes; D009124:Muscle Proteins; C085834:PA700 proteasome activator; C085047:PSME1 protein, human; D011506:Proteins; D007371:Interferon-gamma; D010447:Peptide Hydrolases; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease; D000251:Adenosine Triphosphatases