Separation of yeast proteasome subunits. Immunoreactivity with antibodies against ATP-dependent protease Ti from Escherichia coli.
On two-dimensional gel electrophoresis, proteasomes (multicatalytic proteinase complexes) from the yeast Saccharomyces cerevisiae were separated into a characteristic set of approximately 20 components with molecular weights of 21,000 to 31,000 and isoelectric points of 3.5 to 7.5. The main components were isolated by reverse-phase high performance liquid chromatography on a TSK gel phenyl-5PW RP column and named YC1 to YC11, in order of their elution. Immuno-blot analysis showed that two components (YC1-alpha and YC1-beta) with molecular weights of 30,800 and 28,300 strongly cross-reacted with antibody against the P-component of ATP-dependent protease Ti from Escherichia coli, but no components were found to react with antibodies against the A-component of protease Ti or another ATP-dependent protease La (the Ion gene product) of Escherichia coli. These results indicate a structural relationship between eukaryotic proteasomes and bacterial ATP-dependent protease Ti.
Tanaka
K
K
Institute for Enzyme Research, University of Tokushima, Japan.
Tamura
T
T
Kumatori
A
A
Kwak
T H
TH
Chung
C H
CH
Ichihara
A
A
D000251:Adenosine Triphosphatases; D000906:Antibodies; D000936:Antigen-Antibody Complex; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D003546:Cysteine Endopeptidases / Q000276:immunology / Q000302:isolation & purification*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D049071:Endopeptidase Clp; D004926:Escherichia coli / Q000201:enzymology; D015151:Immunoblotting; D046911:Macromolecular Substances; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000276:immunology / Q000302:isolation & purification*; D046988:Proteasome Endopeptidase Complex; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D012697:Serine Endopeptidases / Q000276:immunology / Q000302:isolation & purification*
D000906:Antibodies; D000936:Antigen-Antibody Complex; D046911:Macromolecular Substances; D009097:Multienzyme Complexes; D012697:Serine Endopeptidases; D049071:Endopeptidase Clp; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; D000251:Adenosine Triphosphatases