Characterizing the specificity and cooperation of aminopeptidases in the cytosol and endoplasmic reticulum during MHC class I antigen presentation.
Many MHC class I-binding peptides are generated as N-extended precursors during protein degradation by the proteasome. These peptides can subsequently be trimmed by aminopeptidases in the cytosol and/or the endoplasmic reticulum (ER) to produce mature epitope. However, the contribution and specificity of each of these subcellular compartments in removing N-terminal amino acids for Ag presentation is not well defined. In this study, we investigated this issue for antigenic precursors that are expressed in the cytosol. By systematically varying the N-terminal flanking sequences of peptides, we show that the amino acids upstream of an epitope precursor are a major determinant of the amount of Ag presentation. In many cases, MHC class I-binding peptides are produced through sequential trimming in the cytosol and ER. Trimming of flanking residues in the cytosol contributes most to sequences that are poorly trimmed in the ER. Because N-terminal trimming has different specificity in the cytosol and ER, the cleavage of peptides in both of these compartments serves to broaden the repertoire of sequences that are presented.
Hearn
Arron
A
Department of Pathology, University of Massachusetts Medical School, Worcester, MA 01655, USA.
York
Ian A
IA
Bishop
Courtney
C
Rock
Kenneth L
KL
D000626:Aminopeptidases / Q000502:physiology*; D000818:Animals; D017951:Antigen Presentation / Q000276:immunology*; D003600:Cytosol / Q000201:enzymology* / Q000276:immunology* / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000201:enzymology* / Q000276:immunology* / Q000378:metabolism; D018984:Epitopes, T-Lymphocyte / Q000276:immunology / Q000378:metabolism; D006183:H-2 Antigens / Q000276:immunology* / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010446:Peptide Fragments / Q000276:immunology / Q000378:metabolism; D011498:Protein Precursors / Q000276:immunology / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000276:immunology; D013379:Substrate Specificity / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
D018984:Epitopes, T-Lymphocyte; D006183:H-2 Antigens; C032876:H-2Kb protein, mouse; D010446:Peptide Fragments; D011498:Protein Precursors; D000626:Aminopeptidases