{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T10:30:01Z","timestamp":1789727401712,"version":"4.0.1"},"reference-count":45,"publisher":"Oxford University Press (OUP)","issue":"8","license":[{"start":{"date-parts":[[2010,10,1]],"date-time":"2010-10-01T00:00:00Z","timestamp":1285891200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2010,10,15]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>As a type II transmembrane protein in basal keratinocytes, collagen XVII provides stable adhesion between epidermis and dermis in the skin. Its ectodomain can be shed from the cell surface, and autoantibodies in certain blistering diseases preferentially recognize the shed form. Major epitopes of collagen XVII are clustered within the juxtamembranous noncollagenous 16th A domain, and ectodomain shedding occurs within this region, suggesting that cleavage generates neoepitopes. However, the candidate cleavage sites have been controversial, and the mechanism of neoepitope generation is unclear. In this study, we investigated cleavage sites in the noncollagenous 16th A domain to understand the generation of neoepitopes and their pathological role. Polyclonal Abs recognizing the stretch Leu524-Gly532 preferentially reacted with the shed ectodomain, but not with the full-length form, indicating that a neoepitope was localized at this site. The neoepitope-specific Ab fixed complement and induced granulocyte-dependent dermal-epidermal separation in cryosections of normal human skin. The physiological cleavage sites were identified using mass spectrometry. N termini were found at Asp514, Leu524, Glu525, and Gly526, among which Asp514 and Glu525 were blocked by acetylation and pyroglutaminate. In silico prediction of B cell epitopes indicated that the antigenicity of the Leu524-Gly532 region increased substantially after shedding, regardless of the cleavage sites. Correspondingly, neoepitopes were found in the skin and blister fluids of patients with bullous pemphigoid, and bullous pemphigoid sera reacted with the peptide Leu524-Gly532. Taken together, these data demonstrate that physiological shedding of collagen XVII generates neoepitopes, which may serve as a target of blister-inducing autoantibodies.<\/jats:p>","DOI":"10.4049\/jimmunol.1001524","type":"journal-article","created":{"date-parts":[[2010,9,23]],"date-time":"2010-09-23T00:40:12Z","timestamp":1285202412000},"page":"4938-4947","source":"Crossref","is-referenced-by-count":65,"title":["Ectodomain Shedding Generates Neoepitopes on Collagen XVII, the Major Autoantigen for Bullous Pemphigoid"],"prefix":"10.1093","volume":"185","author":[{"given":"Wataru","family":"Nishie","sequence":"first","affiliation":[{"name":"Department of Dermatology, University Medical Center Freiburg , Freiburg ,","place":["Germany"]},{"name":"Department of Dermatology, Hokkaido University Graduate School of Medicine , Sapporo ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephanie","family":"Lamer","sequence":"additional","affiliation":[{"name":"Core Facility Proteomics, Center for Systems Biology (ZBSA) , Freiburg ,","place":["Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas","family":"Schlosser","sequence":"additional","affiliation":[{"name":"Core Facility Proteomics, Center for Systems Biology (ZBSA) , Freiburg ,","place":["Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emilia","family":"Licarete","sequence":"additional","affiliation":[{"name":"Department of Dermatology, University Medical Center Freiburg , Freiburg ,","place":["Germany"]},{"name":"Department of Biology, Babes-Bolyai University of Cluj-Napoca , Cluj-Napoca ,","place":["Romania"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claus-Werner","family":"Franzke","sequence":"additional","affiliation":[{"name":"Department of Dermatology, University Medical Center Freiburg , Freiburg ,","place":["Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silke C","family":"Hofmann","sequence":"additional","affiliation":[{"name":"Department of Dermatology, University Medical Center Freiburg , Freiburg ,","place":["Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joanna","family":"Jackow","sequence":"additional","affiliation":[{"name":"Department of Dermatology, University Medical Center Freiburg , Freiburg ,","place":["Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cassian","family":"Sitaru","sequence":"additional","affiliation":[{"name":"Department of Dermatology, University Medical Center Freiburg , Freiburg ,","place":["Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leena","family":"Bruckner-Tuderman","sequence":"additional","affiliation":[{"name":"Department of Dermatology, University Medical Center Freiburg , Freiburg ,","place":["Germany"]},{"name":"Freiburg Institute for Advanced Studies, School of Life Sciences-LifeNet, University of Freiburg , Freiburg ,","place":["Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2010,10,15]]},"reference":[{"key":"2025032412002141600_r1","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1038\/nrm2120","article-title":"In search of partners: linking extracellular proteases to substrates","volume":"8","author":"Overall","year":"2007","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"2025032412002141600_r2","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1016\/j.semcdb.2008.11.002","article-title":"The \u201ca disintegrin and metalloprotease\u201d (ADAM) family of sheddases: physiological and cellular functions","volume":"20","author":"Reiss","year":"2009","journal-title":"Semin. Cell Dev. Biol."},{"key":"2025032412002141600_r3","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1016\/j.tibs.2005.05.006","article-title":"Shedding light on ADAM metalloproteinases","volume":"30","author":"Huovila","year":"2005","journal-title":"Trends Biochem. Sci."},{"key":"2025032412002141600_r4","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1016\/S0945-053X(03)00051-9","article-title":"Collagenous transmembrane proteins: collagen XVII as a prototype","volume":"22","author":"Franzke","year":"2003","journal-title":"Matrix Biol."},{"key":"2025032412002141600_r5","doi-asserted-by":"crossref","first-page":"4005","DOI":"10.1074\/jbc.R400034200","article-title":"Collagenous transmembrane proteins: recent insights into biology and pathology","volume":"280","author":"Franzke","year":"2005","journal-title":"J. Biol. Chem."},{"key":"2025032412002141600_r6","doi-asserted-by":"crossref","first-page":"8825","DOI":"10.1016\/S0021-9258(18)52948-3","article-title":"Cloning of type XVII collagen: complementary and genomic DNA sequences of mouse 180-kilodalton bullous pemphigoid antigen (BPAG2) predict an interrupted collagenous domain, a transmembrane segment, and unusual features in the 5\u2032-end of the gene and the 3\u2032-untranslated region of the mRNA","volume":"268","author":"Li","year":"1993","journal-title":"J. Biol. Chem."},{"key":"2025032412002141600_r7","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1111\/j.1365-2133.1995.tb02665.x","article-title":"Expression pattern of the bullous pemphigoid-180 antigen in normal and neoplastic epithelia","volume":"133","author":"Fairley","year":"1995","journal-title":"Br. J. Dermatol."},{"key":"2025032412002141600_r8","first-page":"352","article-title":"Cloning of the human type XVII collagen gene (COL17A1), and detection of novel mutations in generalized atrophic benign epidermolysis bullosa","volume":"60","author":"Gatalica","year":"1997","journal-title":"Am. J. Hum. Genet."},{"key":"2025032412002141600_r9","doi-asserted-by":"crossref","first-page":"1560","DOI":"10.1073\/pnas.88.4.1560","article-title":"cDNA analysis predicts a cornea-specific collagen","volume":"88","author":"Marchant","year":"1991","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2025032412002141600_r10","doi-asserted-by":"crossref","first-page":"91","DOI":"10.2353\/ajpath.2009.080573","article-title":"Type XVII collagen is a key player in tooth enamel formation","volume":"174","author":"Asaka","year":"2009","journal-title":"Am. J. Pathol."},{"key":"2025032412002141600_r11","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/j.matbio.2005.11.004","article-title":"Collagen XVII is expressed in human CNS neurons","volume":"25","author":"Sepp\u00e4nen","year":"2006","journal-title":"Matrix Biol."},{"key":"2025032412002141600_r12","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1038\/ng0995-83","article-title":"Mutations in the 180-kD bullous pemphigoid antigen (BPAG2), a hemidesmosomal transmembrane collagen (COL17A1), in generalized atrophic benign epidermolysis bullosa","volume":"11","author":"McGrath","year":"1995","journal-title":"Nat. Genet."},{"key":"2025032412002141600_r13","doi-asserted-by":"crossref","first-page":"25937","DOI":"10.1074\/jbc.273.40.25937","article-title":"Two forms of collagen XVII in keratinocytes: a full-length transmembrane protein and a soluble ectodomain","volume":"273","author":"Sch\u00e4cke","year":"1998","journal-title":"J. Biol. Chem."},{"key":"2025032412002141600_r14","doi-asserted-by":"crossref","first-page":"9711","DOI":"10.1074\/jbc.273.16.9711","article-title":"Cleavage of BP180, a 180-kDa bullous pemphigoid antigen, yields a 120-kDa collagenous extracellular polypeptide","volume":"273","author":"Hirako","year":"1998","journal-title":"J. Biol. Chem."},{"key":"2025032412002141600_r15","doi-asserted-by":"crossref","first-page":"5026","DOI":"10.1093\/emboj\/cdf532","article-title":"Transmembrane collagen XVII, an epithelial adhesion protein, is shed from the cell surface by ADAMs","volume":"21","author":"Franzke","year":"2002","journal-title":"EMBO J."},{"key":"2025032412002141600_r16","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1093\/jb\/mvg024","article-title":"Extracellular cleavage of bullous pemphigoid antigen 180\/type XVII collagen and its involvement in hemidesmosomal disassembly","volume":"133","author":"Hirako","year":"2003","journal-title":"J. Biochem."},{"key":"2025032412002141600_r17","doi-asserted-by":"crossref","first-page":"24521","DOI":"10.1074\/jbc.M308835200","article-title":"Shedding of collagen XVII\/BP180: structural motifs influence cleavage from cell surface","volume":"279","author":"Franzke","year":"2004","journal-title":"J. Biol. Chem."},{"key":"2025032412002141600_r18","doi-asserted-by":"crossref","first-page":"23386","DOI":"10.1074\/jbc.M109.034090","article-title":"Shedding of collagen XVII\/BP180 in skin depends on both ADAM10 and ADAM9","volume":"284","author":"Franzke","year":"2009","journal-title":"J. Biol. Chem."},{"key":"2025032412002141600_r19","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1046\/j.1523-1747.1998.00129.x","article-title":"The 97 kDa linear IgA bullous disease antigen is identical to a portion of the extracellular domain of the 180 kDa bullous pemphigoid antigen, BPAg2","volume":"110","author":"Zone","year":"1998","journal-title":"J. Invest. Dermatol."},{"key":"2025032412002141600_r20","doi-asserted-by":"crossref","first-page":"1554","DOI":"10.1046\/j.1523-1747.2003.12607.x","article-title":"The 97-kDa (LABD97) and 120-kDa (LAD-1) fragments of bullous pemphigoid antigen 180\/type XVII collagen have different N-termini","volume":"121","author":"Hirako","year":"2003","journal-title":"J. Invest. Dermatol."},{"key":"2025032412002141600_r21","doi-asserted-by":"crossref","first-page":"22737","DOI":"10.1074\/jbc.M701937200","article-title":"Extracellular phosphorylation of collagen XVII by ecto-casein kinase 2 inhibits ectodomain shedding","volume":"282","author":"Zimina","year":"2007","journal-title":"J. Biol. Chem."},{"key":"2025032412002141600_r22","doi-asserted-by":"crossref","first-page":"1730","DOI":"10.1038\/jid.2008.424","article-title":"Plasmin plays a role in the in vitro generation of the linear IgA dermatosis antigen LADB97","volume":"129","author":"Hofmann","year":"2009","journal-title":"J. Invest. Dermatol."},{"key":"2025032412002141600_r23","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1136\/mp.52.1.11","article-title":"The use of cleavage site specific antibodies to delineate protein processing and breakdown pathways","volume":"52","author":"Mort","year":"1999","journal-title":"Mol. Pathol."},{"key":"2025032412002141600_r24","doi-asserted-by":"crossref","first-page":"3512","DOI":"10.4049\/jimmunol.0803573","article-title":"Autoantibodies against complement C1q specifically target C1q bound on early apoptotic cells","volume":"183","author":"Bigler","year":"2009","journal-title":"J. Immunol."},{"key":"2025032412002141600_r25","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1016\/S0002-9440(10)64772-4","article-title":"The shed ectodomain of collagen XVII\/BP180 is targeted by autoantibodies in different blistering skin diseases","volume":"156","author":"Schumann","year":"2000","journal-title":"Am. J. Pathol."},{"key":"2025032412002141600_r26","doi-asserted-by":"crossref","first-page":"734","DOI":"10.1111\/1523-1747.ep12345782","article-title":"LAD-1, the linear IgA bullous dermatosis autoantigen, is a novel 120-kDa anchoring filament protein synthesized by epidermal cells","volume":"106","author":"Marinkovich","year":"1996","journal-title":"J. Invest. Dermatol."},{"key":"2025032412002141600_r27","doi-asserted-by":"crossref","first-page":"757","DOI":"10.1097\/01.AOG.0000115506.76104.ad","article-title":"Immunoblotting and enzyme-linked immunosorbent assay for the diagnosis of pemphigoid gestationis","volume":"103","author":"Sitaru","year":"2004","journal-title":"Obstet. Gynecol."},{"key":"2025032412002141600_r28","doi-asserted-by":"crossref","first-page":"664","DOI":"10.1046\/j.1523-1747.2002.01720.x","article-title":"Autoantibodies to bullous pemphigoid antigen 180 induce dermal-epidermal separation in cryosections of human skin","volume":"118","author":"Sitaru","year":"2002","journal-title":"J. Invest. Dermatol."},{"key":"2025032412002141600_r29","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1093\/protein\/gzn075","article-title":"COBEpro: a novel system for predicting continuous B-cell epitopes","volume":"22","author":"Sweredoski","year":"2009","journal-title":"Protein Eng. Des. Sel."},{"key":"2025032412002141600_r30","doi-asserted-by":"crossref","first-page":"9526","DOI":"10.1021\/ac801708p","article-title":"Evaluation of the low-specificity protease elastase for large-scale phosphoproteome analysis","volume":"80","author":"Wang","year":"2008","journal-title":"Anal. Chem."},{"key":"2025032412002141600_r31","doi-asserted-by":"crossref","first-page":"30631","DOI":"10.1074\/jbc.M103748200","article-title":"Regulation of membrane metalloproteolytic cleavage of L-selectin (CD62l) by the epidermal growth factor domain","volume":"276","author":"Zhao","year":"2001","journal-title":"J. Biol. Chem."},{"key":"2025032412002141600_r32","doi-asserted-by":"crossref","first-page":"1164","DOI":"10.1046\/j.1523-1747.2000.0202a-7.x","article-title":"IgA antibodies of linear IgA bullous dermatosis recognize the 15th collagenous domain of BP180","volume":"115","author":"Nie","year":"2000","journal-title":"J. Invest. Dermatol."},{"key":"2025032412002141600_r33","doi-asserted-by":"crossref","first-page":"924","DOI":"10.1038\/jid.2010.14","article-title":"How does intramolecular epitope spreading occur in BPAG2 (BP180)?","volume":"130","author":"Hashimoto","year":"2010","journal-title":"J. Invest. Dermatol."},{"key":"2025032412002141600_r34","doi-asserted-by":"crossref","first-page":"947","DOI":"10.1046\/j.1523-1747.1999.00808.x","article-title":"Autoantibodies in a subgroup of patients with linear IgA disease react with the NC16A domain of BP1801","volume":"113","author":"Zillikens","year":"1999","journal-title":"J. Invest. Dermatol."},{"key":"2025032412002141600_r35","doi-asserted-by":"crossref","first-page":"1032","DOI":"10.1016\/j.bbrc.2006.09.147","article-title":"The NC16A domain of collagen XVII plays a role in triple helix assembly and stability","volume":"350","author":"Van den Bergh","year":"2006","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"2025032412002141600_r36","doi-asserted-by":"crossref","first-page":"3093","DOI":"10.1074\/jbc.275.5.3093","article-title":"Collagen XVII is destabilized by a glycine substitution mutation in the cell adhesion domain Col15","volume":"275","author":"Tasanen","year":"2000","journal-title":"J. Biol. Chem."},{"key":"2025032412002141600_r37","doi-asserted-by":"crossref","first-page":"986","DOI":"10.1046\/j.1365-2133.2001.04528.x","article-title":"False-negative results in immunoblot assay of serum IgA antibodies reactive with the 180-kDa bullous pemphigoid antigen: the importance of primary incubation temperature","volume":"145","author":"Pas","year":"2001","journal-title":"Br. J. Dermatol."},{"key":"2025032412002141600_r38","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1002\/path.1674","article-title":"Granulocyte-derived elastase and gelatinase B are required for dermal-epidermal separation induced by autoantibodies from patients with epidermolysis bullosa acquisita and bullous pemphigoid","volume":"204","author":"Shimanovich","year":"2004","journal-title":"J. Pathol."},{"key":"2025032412002141600_r39","doi-asserted-by":"crossref","first-page":"2480","DOI":"10.1172\/JCI116856","article-title":"A passive transfer model of the organ-specific autoimmune disease, bullous pemphigoid, using antibodies generated against the hemidesmosomal antigen, BP180","volume":"92","author":"Liu","year":"1993","journal-title":"J. Clin. Invest."},{"key":"2025032412002141600_r40","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1038\/nm1496","article-title":"Humanization of autoantigen","volume":"13","author":"Nishie","year":"2007","journal-title":"Nat. Med."},{"key":"2025032412002141600_r41","doi-asserted-by":"crossref","first-page":"4088","DOI":"10.4049\/jimmunol.0800389","article-title":"A novel humanized neonatal autoimmune blistering skin disease model induced by maternally transferred antibodies","volume":"183","author":"Nishie","year":"2009","journal-title":"J. Immunol."},{"key":"2025032412002141600_r42","doi-asserted-by":"crossref","first-page":"955","DOI":"10.1038\/nmeth811","article-title":"Positional proteomics: selective recovery and analysis of N-terminal proteolytic peptides","volume":"2","author":"McDonald","year":"2005","journal-title":"Nat. Methods"},{"key":"2025032412002141600_r43","doi-asserted-by":"crossref","first-page":"1790","DOI":"10.1038\/nprot.2006.317","article-title":"Positional proteomics: preparation of amino-terminal peptides as a strategy for proteome simplification and characterization","volume":"1","author":"McDonald","year":"2006","journal-title":"Nat. Protoc."},{"key":"2025032412002141600_r44","doi-asserted-by":"crossref","first-page":"978","DOI":"10.1172\/JCI116674","article-title":"The autoimmune blistering skin disease bullous pemphigoid: the presence of plasmin\/\u03b1 2-antiplasmin complexes in skin blister fluid indicates plasmin generation in lesional skin","volume":"92","author":"Kramer","year":"1993","journal-title":"J. Clin. Invest."},{"key":"2025032412002141600_r45","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1046\/j.1365-2133.1998.02028.x","article-title":"Internalization of the 180 kDa bullous pemphigoid antigen as immune complexes in basal keratinocytes: an important early event in blister formation in bullous pemphigoid","volume":"138","author":"Kitajima","year":"1998","journal-title":"Br. J. Dermatol."}],"container-title":["The Journal of Immunology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/jimmunol\/article-pdf\/185\/8\/4938\/61612677\/1001524.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/jimmunol\/article-pdf\/185\/8\/4938\/61612677\/1001524.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,3,28]],"date-time":"2025-03-28T21:24:51Z","timestamp":1743197091000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/jimmunol\/article\/185\/8\/4938\/7977864"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2010,10]]},"references-count":45,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2010,10,15]]}},"URL":"https:\/\/doi.org\/10.4049\/jimmunol.1001524","relation":{"has-review":[{"id-type":"doi","id":"10.3410\/f.6169956.10176054","asserted-by":"object"},{"id-type":"doi","id":"10.3410\/f.6169956.6214054","asserted-by":"object"}]},"ISSN":["0022-1767","1550-6606"],"issn-type":[{"value":"0022-1767","type":"print"},{"value":"1550-6606","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2010,10]]},"published":{"date-parts":[[2010,10]]}}}