Adrian Martín-Esteban1, Pablo Guasp1, Eilon Barnea2, Arie Admon2, José A López de Castro3. 1. Centro de Biología Molecular Severo Ochoa (Consejo Superior de Investigaciones Científicas and Universidad Autónoma), Madrid, Spain. 2. Technion-Israel Institute of Technology, Haifa, Israel. 3. Centro de Biología Molecular Severo Ochoa (Consejo Superior de Investigaciones Científicas and Universidad Autónoma), Madrid, Spain. aldecastro@cbm.csic.es.
Abstract
OBJECTIVE: To determine the influence of endoplasmic reticulum aminopeptidase 2 (ERAP-2) expression on the HLA-B*27 peptidome in live cells. METHODS: Using immunoaffinity chromatography and acid extraction, HLA-B*27:05-bound peptides were isolated from 2 ERAP-2-negative lymphoblastoid cell lines and 1 ERAP-2-positive lymphoblastoid cell line expressing functionally indistinguishable ERAP-1 variants. More than 2,000-4,000 B*27:05 ligands were identified from each cell line, and their relative abundance was established by quantitative tandem mass spectrometry and MaxQuant-based peptide analyses. Pairwise comparisons were used to determine the structural features of peptides whose relative abundance was dependent on the presence of ERAP-2. Synthetic peptide digestions were performed with recombinant ERAP-1 and ERAP-2. Peptide affinity was estimated with standard algorithms. RESULTS: The B*27:05 peptidome from ERAP-2-positive cells showed 3-4% fewer peptides with N-terminal basic residues than did the peptidome from ERAP-2-negative cells. Among the shared peptides, those most abundant in the presence of ERAP-2 included more nonamers, fewer decamers, and fewer N-terminal basic residues than the peptides predominant in ERAP-2-negative cells. These ERAP-2-dependent changes did not alter the global affinity of the B*27:05 peptidome. CONCLUSION: ERAP-2 significantly influences the B*27:05-bound peptidome by destroying some ligands and decreasing the abundance of many more ligands with N-terminal basic residues, while increasing the abundance of nonamers. The former effects are best explained by direct ERAP-2 trimming. The effects on peptide length might be attributed to ERAP-2-induced activation of ERAP-1 trimming. These data support the notion of a peptide-mediated mechanism as the basis for the association of ERAP-2 with ankylosing spondylitis. Analogous effects on other major histocompatibility complex class I peptidomes might explain the involvement of ERAP-2 in HLA-B27-negative spondyloarthritis.
OBJECTIVE: To determine the influence of endoplasmic reticulum aminopeptidase 2 (ERAP-2) expression on the HLA-B*27 peptidome in live cells. METHODS: Using immunoaffinity chromatography and acid extraction, HLA-B*27:05-bound peptides were isolated from 2 ERAP-2-negative lymphoblastoid cell lines and 1 ERAP-2-positive lymphoblastoid cell line expressing functionally indistinguishable ERAP-1 variants. More than 2,000-4,000 B*27:05 ligands were identified from each cell line, and their relative abundance was established by quantitative tandem mass spectrometry and MaxQuant-based peptide analyses. Pairwise comparisons were used to determine the structural features of peptides whose relative abundance was dependent on the presence of ERAP-2. Synthetic peptide digestions were performed with recombinant ERAP-1 and ERAP-2. Peptide affinity was estimated with standard algorithms. RESULTS: The B*27:05 peptidome from ERAP-2-positive cells showed 3-4% fewer peptides with N-terminal basic residues than did the peptidome from ERAP-2-negative cells. Among the shared peptides, those most abundant in the presence of ERAP-2 included more nonamers, fewer decamers, and fewer N-terminal basic residues than the peptides predominant in ERAP-2-negative cells. These ERAP-2-dependent changes did not alter the global affinity of the B*27:05 peptidome. CONCLUSION:ERAP-2 significantly influences the B*27:05-bound peptidome by destroying some ligands and decreasing the abundance of many more ligands with N-terminal basic residues, while increasing the abundance of nonamers. The former effects are best explained by direct ERAP-2 trimming. The effects on peptide length might be attributed to ERAP-2-induced activation of ERAP-1 trimming. These data support the notion of a peptide-mediated mechanism as the basis for the association of ERAP-2 with ankylosing spondylitis. Analogous effects on other major histocompatibility complex class I peptidomes might explain the involvement of ERAP-2 in HLA-B27-negative spondyloarthritis.
Authors: Pablo Guasp; Elena Lorente; Adrian Martín-Esteban; Eilon Barnea; Paolo Romania; Doriana Fruci; JonasJ W Kuiper; Arie Admon; José A López de Castro Journal: Mol Cell Proteomics Date: 2019-05-15 Impact factor: 5.911
Authors: Vidya Ranganathan; Eric Gracey; Matthew A Brown; Robert D Inman; Nigil Haroon Journal: Nat Rev Rheumatol Date: 2017-04-27 Impact factor: 20.543
Authors: Pablo Guasp; Eilon Barnea; M Francisca González-Escribano; Anaïs Jiménez-Reinoso; José R Regueiro; Arie Admon; José A López de Castro Journal: J Biol Chem Date: 2017-04-26 Impact factor: 5.157
Authors: Elena Lorente; Jennifer Redondo-Antón; Adrian Martín-Esteban; Pablo Guasp; Eilon Barnea; Pilar Lauzurica; Arie Admon; José A López de Castro Journal: Mol Cell Proteomics Date: 2019-09-17 Impact factor: 5.911
Authors: Alejandro Sanz-Bravo; Adrian Martín-Esteban; Jonas J W Kuiper; Marina García-Peydró; Eilon Barnea; Arie Admon; José A López de Castro Journal: Mol Cell Proteomics Date: 2018-05-16 Impact factor: 5.911
Authors: Elena Lorente; Miguel G Fontela; Eilon Barnea; Antonio J Martín-Galiano; Carmen Mir; Begoña Galocha; Arie Admon; Pilar Lauzurica; Daniel López Journal: Mol Cell Proteomics Date: 2020-04-07 Impact factor: 5.911