Different proteasome subtypes in a single tissue exhibit different enzymatic properties.
It is concluded from many experiments that mammalian tissues and cells must contain a heterogeneous population of 20 S proteasome complexes. We describe the purification and separation by chromatographic procedures of constitutive 20 S proteasomes, 20 S immuno-proteasomes and intermediate-type 20 S proteasomes from a given tissue. Our data demonstrate that each of these three groups comprises more than one subtype and that the relative ratios of the subtypes differ between different rat tissues. Thus, six subtypes could be identified in rat muscle tissue. Subtypes I and II are constitutive proteasomes, while subtypes V and VI comprise immuno-proteasomes. Subtypes III and IV belong to a group of intermediate-type proteasomes. The subtypes differ with regard to their enzymatic characteristics. Subtypes I-III exhibit high chymotrypsin-like activity and high peptidylglutamylpeptide hydrolysing activity, while these activities are depressed in subtypes IV-VI. In contrast, trypsin-like activity of subtypes IV-VI is enhanced in comparison to subtypes I-III. Importantly, the subtypes also differ in their preferential cleavage site usage when tested by digestion of a synthetic 25mer polypeptide substrate. Therefore, the characteristics of proteasomes purified from tissues or cells represent the average of the different subtype activities which in turn may have different functions in vivo.
Dahlmann
B
B
Department of Clinical Biochemistry, Deutsches Diabetes-Forschungsinstitut, Düsseldorf, Germany. Burkhardt.Dahlmann@charite.de
Ruppert
T
T
Kuehn
L
L
Merforth
S
S
Kloetzel
P M
PM
D000595:Amino Acid Sequence; D000818:Animals; D003546:Cysteine Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D017874:Immediate-Early Proteins / Q000737:chemistry / Q000378:metabolism; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008297:Male; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000201:enzymology*; D009928:Organ Specificity; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex; D021122:Protein Subunits; D051381:Rats; D017208:Rats, Wistar; D013379:Substrate Specificity
D017874:Immediate-Early Proteins; D007527:Isoenzymes; D009097:Multienzyme Complexes; D010446:Peptide Fragments; D021122:Protein Subunits; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex