Abstract
The generation of many HLA class I peptides entails a final trimming step in the endoplasmic reticulum that, in humans, is accomplished by two 'candidate' aminopeptidases. We show here that one of these, ERAP1, was unable to remove several N-terminal amino acids that were trimmed efficiently by the second enzyme, ERAP2. This trimming of a longer peptide required the concerted action of both ERAP1 and ERAP2, both for in vitro digestion and in vivo for cellular antigen presentation. ERAP1 and ERAP2 localized together in vivo and associated physically in complexes that were most likely heterodimeric. Thus, the human endoplasmic reticulum is equipped with a pair of trimming aminopeptidases that have complementary functions in HLA class I peptide presentation.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on SpringerLink
- Instant access to the full article PDF.
USD 39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
The ERAP1 active site cannot productively access the N-terminus of antigenic peptide precursors stably bound onto MHC class I
Conformational dynamics linked to domain closure and substrate binding explain the ERAP1 allosteric regulation mechanism
Evolutionary immuno-genetics of endoplasmic reticulum aminopeptidase II (ERAP2)
References
Shastri, N., Schwab, S. & Serwold, T. Producing nature's gene-chips: the generation of peptides for display by MHC class I molecules. Annu. Rev. Immunol. 20, 463–493 (2002).
Rock, K.L. & Goldberg, A.L. Degradation of cell proteins and the generation of MHC class I- presented peptides. Annu. Rev. Immunol. 17, 739–779 (1999).
Reits, E. et al. A major role for TPPII in trimming proteasomal degradation products for MHC class I antigen presentation. Immunity 20, 495–506 (2004).
Neisig, A. et al. Major differences in transporter associated with antigen presentation (TAP)-dependent translocation of MHC clas I-presentable peptides and the effect of flanking sequences. J. Immunol. 154, 1273–1279 (1995).
Daniel, S. et al. Relationship between peptide selectivities of human transporters associated with antigen processing and HLA class I molecules. J. Immunol. 161, 617–624 (1998).
Lauvau, G. et al. Human transporters associated with antigen processing (TAPs) select epitope precursor peptides for processing in the endoplasmic reticulum and presentation to T cells. J. Exp. Med. 190, 1227–1240 (1999).
Cui, X. et al. Identification of ARTS-1 as a novel TNFR1-binding protein that promotes TNFR1 ectodomain shedding. J. Clin. Invest. 110, 515–526 (2002).
Hattori, A., Matsumoto, H., Mizutani, S. & Tsujimoto, M. Molecular cloning of adipocyte-derived leucine aminopeptidase highly related to placental leucine aminopeptidase/oxytocinase. J. Biochem. 125, 931–938 (1999).
Schomburg, L., Kollmus, H., Friedrichsen, S. & Bauer, K. Molecular characterization of a puromycin-insensitive leucyl-specific aminopeptidase, PILS-AP. Eur. J. Biochem. 267, 3198–3207 (2000).
Serwold, T., Gonzalez, F., Kim, J., Jacob, R. & Shastri, N. ERAAP customizes peptides for MHC class I molecules in the endoplasmic reticulum. Nature 419, 480–483 (2002).
York, I.A. et al. The ER aminopeptidase ERAP1 enhances or limits antigen presentation by trimming epitopes to 8–9 residues. Nat. Immunol. 3, 1177–1184 (2002).
Saric, T. et al. An IFN-γ-induced aminopeptidase in the ER, ERAP1, trims precursors to MHC class I-presented peptides. Nat. Immunol. 3, 1169–1176 (2002).
Serwold, T., Gaw, S. & Shastri, N. ER aminopeptidases generate a unique pool of peptides for MHC class I molecules. Nat. Immunol. 2, 644–651 (2001).
Tanioka, T. et al. Human leukocyte-derived arginine aminopeptidase. The third member of the oxytocinase subfamily of aminopeptidases. J. Biol. Chem. 278, 32275–32283 (2003).
Fruci, D., Niedermann, G., Butler, R.H. & van Endert, P.M. Efficient MHC class I-independent amino-terminal trimming of epitope precursor peptides in the endoplasmic reticulum. Immunity 15, 467–476 (2001).
Rowe, M. et al. Restoration of endogenous antigen processing in Burkitt's lymphoma cells by Epstein-Barr virus latent membrane protein-1: coordinate up-regulation of peptide transporters and HLA-class I antigen expression. Eur. J. Immunol. 25, 1374–1384 (1995).
Samino, Y., Lopez, D., Guil, S., de Leon, P. & Del Val, M. An endogenous HIV envelope-derived peptide without the terminal NH3 + group anchor is physiologically presented by major histocompatibility complex class I molecules. J. Biol. Chem. 279, 1151–1160 (2004).
Roelse, J., Grommé, M., Momburg, F., Hämmerling, G. & Neefjes, J. Trimming of TAP-translocated peptides in the endoplasmic reticulum and in the cytosol during recycling. J. Exp. Med. 180, 1591–1597 (1994).
Koopmann, J.O. et al. Export of antigenic peptides from the endoplasmic reticulum intersects with retrograde protein translocation through the Sec61p channel. Immunity 13, 117–127 (2000).
Früh, K. et al. A viral inhibitor of peptide transporters for antigen presentation. Nature 375, 415–418 (1995).
Brouwenstijn, N., Serwold, T. & Shastri, N. MHC class I molecules can direct proteolytic cleavage of antigenic precursors in the endoplasmic reticulum. Immunity 15, 95–104 (2001).
Gibbs, R.A. et al. Genome sequence of the Brown Norway rat yields insights into mammalian evolution. Nature 428, 493–521 (2004).
Matsumoto, H. et al. Characterization of a recombinant soluble form of human placental leucine aminopeptidase/oxytocinase expressed in Chinese hamster ovary cells. Eur. J. Biochem. 267, 46–52 (2000).
Rock, K.L., York, I.A. & Goldberg, A.L. Post-proteasomal antigen processing for major histocompatibility complex class I presentation. Nat. Immunol. 5, 670–677 (2004).
Van Endert, P.M. et al. The peptide-binding motif for the human transporter associated with antigen processing. J. Exp. Med. 182, 1883–1895 (1995).
Uebel, S. et al. Recognition principle of the TAP transporter disclosed by combinatorial peptide libraries. Proc. Natl. Acad. Sci. USA 94, 8976–8981 (1997).
Hattori, A. et al. Characterization of recombinant human adipocyte-derived leucine aminopeptidase expressed in Chinese hamster ovary cells. J. Biochem. 128, 755–762 (2000).
Saric, T. et al. Major histocompatibility complex class I-presented antigenic peptides are degraded in cytosolic extracts primarily by thimet oligopeptidase. J. Biol. Chem. 276, 36474–36481 (2001).
York, I.A. et al. The cytosolic endopeptidase, thimet oligopeptidase, destroys antigenic peptides and limits the extent of MHC class I antigen presentation. Immunity 18, 429–440 (2003).
van Endert, P.M. et al. A sequential model for peptide binding and transport by the transporters associated with antigen processing. Immunity 1, 491–500 (1994).
Wessel, D. & Flugge, U.I. A method for the quantitative recovery of protein in dilute solutions in the presence of detergents and lipids. Anal. Biochem. 138, 141–143 (1984).
Marks, M.S. Determination of molecular size by zonal sedimentation analysis on sucrose density gradients. in Current Protocols in Cell Biology Vol. 1 (eds. Bonifacino, J.S., Dasso, M., Harford, J.B., Lippincott-Schwartz, J. & Yamada, K.M.) 5.3.1–5.3.33 (John Wiley & Sons, New York, 2000).
Fourneau, J.M., Cohen, H. & van Endert, P.M. A chaperone-assisted high yield system for the production of HLA-DR4 tetramers in insect cells. J. Immunol. Methods 285, 253–264 (2004).
Acknowledgements
We thank Y. Samino for env-specific CTLs and for the purification of env-derived synthetic peptides. Supported by the European Commission (QLK2-CT-2001-01167 to P.M.v.E., F.G. and G.N), Comunidad de Madrid and Fundación para la Investigación y Prevención del Síndrome de Immunodeficiencia Adquirita en Espana (D.L.), Ministerio de Educación y Ciencia and Red Temática de Investigación Cooperativa en Síndrome de Immunodeficiencia Adquirita del Fondo de Investigaciónes Sanitarias (M.D.V.), Fondation pour la Recherche Médicale (D.F.), Institut National de la Santé et de la Recherche Médicale (O.C.) and K. Bauer (MPI Hannover, Germany; L. Schomburg).
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Table 1 (download PDF )
Oligonucleotides used for ERAP1 and ERAP2 knockdown (PDF 28 kb)
Rights and permissions
About this article
Cite this article
Saveanu, L., Carroll, O., Lindo, V. et al. Concerted peptide trimming by human ERAP1 and ERAP2 aminopeptidase complexes in the endoplasmic reticulum. Nat Immunol 6, 689–697 (2005). https://doi.org/10.1038/ni1208
Received:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/ni1208
Share this article
Anyone you share the following link with will be able to read this content:
Sorry, a shareable link is not currently available for this article.
Provided by the Springer Nature SharedIt content-sharing initiative
This article is cited by
-
Polymorphisms in endoplasmic reticulum aminopeptidase genes are associated with cervical cancer risk in a Chinese Han population
BMC Cancer (2020)
-
Plant-derived virus-like particle vaccines drive cross-presentation of influenza A hemagglutinin peptides by human monocyte-derived macrophages
npj Vaccines (2019)
-
Tumor-induced escape mechanisms and their association with resistance to checkpoint inhibitor therapy
Cancer Immunology, Immunotherapy (2019)
-
ERAP1 promotes Hedgehog-dependent tumorigenesis by controlling USP47-mediated degradation of βTrCP
Nature Communications (2019)
-
An allelic variant in the intergenic region between ERAP1 and ERAP2 correlates with an inverse expression of the two genes
Scientific Reports (2018)
