Thimet oligopeptidase and the stability of MHC class I epitopes in macrophage cytosol.
In this study we investigated the fate of a class of proteasome-generated oligopeptides, exposing them to the crude cytosol of macrophages or to the purified recombinant thimet oligopeptidase. Among the proteasome products of known sequences are MHC class I epitopes, 13 of which were randomly chosen to be used as putative substrates. Surprisingly, our results clearly showed that the majority of the peptides were poorly or not degraded, either by the purified enzyme or by the crude macrophage cytosol. The peptides, which were resistant to hydrolysis, displayed high affinity for the thimet oligopeptidase as competitive inhibitors. Regardless of the fact that our data do not allow prediction of whether or not a specific peptide would be degraded, it seems very likely that the structural features, which rule out the stability of the MHC class I peptides in the cytosol, may have implications in an optimized repertoire selection for antigen presentation.
Portaro
F C
FC
Laboratory of Biochemistry and Biophysics, Instituto Butantan, São Paulo, SP, Brazil.
Gomes
M D
MD
Cabrera
A
A
Fernandes
B L
BL
Silva
C L
CL
Ferro
E S
ES
Juliano
L
L
de Camargo
A C
AC
D000818:Animals; D017951:Antigen Presentation / Q000276:immunology; D000938:Antigen-Presenting Cells / Q000201:enzymology; D003546:Cysteine Endopeptidases / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D015395:Histocompatibility Antigens Class I / Q000378:metabolism*; D007700:Kinetics; D008264:Macrophages / Q000201:enzymology*; D008297:Male; D008666:Metalloendopeptidases / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism* / Q000494:pharmacology; D046988:Proteasome Endopeptidase Complex; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity; D013737:Testis / Q000201:enzymology
D004791:Enzyme Inhibitors; D000939:Epitopes; D015395:Histocompatibility Antigens Class I; D009097:Multienzyme Complexes; D009842:Oligopeptides; D011994:Recombinant Proteins; D003546:Cysteine Endopeptidases; D008666:Metalloendopeptidases; C022510:thimet oligopeptidase; D046988:Proteasome Endopeptidase Complex