Discrete cleavage motifs of constitutive and immunoproteasomes revealed by quantitative analysis of cleavage products.
Proteasomes are the main proteases responsible for cytosolic protein degradation and the production of major histocompatibility complex class I ligands. Incorporation of the interferon gamma--inducible subunits low molecular weight protein (LMP)-2, LMP-7, and multicatalytic endopeptidase complex--like (MECL)-1 leads to the formation of immunoproteasomes which have been associated with more efficient class I antigen processing. Although differences in cleavage specificities of constitutive and immunoproteasomes have been observed frequently, cleavage motifs have not been described previously. We now report that cells expressing immunoproteasomes display a different peptide repertoire changing the overall cytotoxic T cell--specificity as indicated by the observation that LMP-7(-/-) mice react against cells of LMP-7 wild-type mice. Moreover, using the 436 amino acid protein enolase-1 as an unmodified model substrate in combination with a quantitative approach, we analyzed a large collection of peptides generated by either set of proteasomes. Inspection of the amino acids flanking proteasomal cleavage sites allowed the description of two different cleavage motifs. These motifs finally explain recent findings describing differential processing of epitopes by constitutive and immunoproteasomes and are important to the understanding of peripheral T cell tolerization/activation as well as for effective vaccine development.
Toes
R E
RE
Institute for Cell Biology, Department of Immunology, University of Tübingen, Germany.
Nussbaum
A K
AK
Degermann
S
S
Schirle
M
M
Emmerich
N P
NP
Kraft
M
M
Laplace
C
C
Zwinderman
A
A
Dick
T P
TP
Müller
J
J
Schönfisch
B
B
Schmid
C
C
Fehling
H J
HJ
Stevanovic
S
S
Rammensee
H G
HG
Schild
H
H
D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D000939:Epitopes; D005260:Female; D008285:Major Histocompatibility Complex; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008817:Mice, Mutant Strains; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D010446:Peptide Fragments / Q000032:analysis / Q000378:metabolism*; D010449:Peptide Mapping; D010751:Phosphopyruvate Hydratase / Q000737:chemistry / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D016038:Skin Transplantation / Q000276:immunology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
D000939:Epitopes; D009097:Multienzyme Complexes; D010446:Peptide Fragments; D011506:Proteins; C071313:LMP-2 protein; D003546:Cysteine Endopeptidases; C077436:LMP7 protein; D046988:Proteasome Endopeptidase Complex; C490920:Psmb10 protein, mouse; D010751:Phosphopyruvate Hydratase