{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T06:50:38Z","timestamp":1791010238843,"version":"4.1.0"},"reference-count":49,"publisher":"National Academy of Sciences","issue":"9","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2006,2,28]]},"abstract":"<jats:p>IL-32, a recently discovered proinflammatory cytokine with four isoforms, induces IL-1\u03b2, TNF-\u03b1, IL-6, and chemokines. Here, we used ligand (IL-32\u03b1) affinity chromatography in an attempt to isolate an IL-32\u03b1 soluble receptor or binding protein. Recombinant IL-32\u03b1 was covalently immobilized on agarose, and preparations of concentrated crude human urinary proteins were applied for chromatographic separation. A specific 30-kDa protein eluted from the column during acid washing and was identified by mass spectrometry as proteinase 3 (PR3) and confirmed by N-terminal microsequencing. PR3, a neutrophil granule serine protease, exists in a soluble or membrane form and is the major autoantigen for autoantibodies in the systemic vasculitic disease, Wegener's granulomatosis. The affinity of IL-32\u03b1 to PR3 was determined by surface plasmon resonance. The dissociation constants were 2.65 \u00b1 0.4 nM for urinary PR3 and 1.2 \u00b1 0.05 nM for neutrophil-derived PR3. However, irreversible inactivation of PR3 enzymatic activity did not significantly change binding to the cytokine. Nevertheless, limited cleavage of IL-32 yielded products consistent with PR3 enzyme activity. Moreover, after limited cleavage by PR3, IL-32\u03b1 was more active than intact IL-32\u03b1 in inducing macrophage inflammatory protein-2 in mouse macrophages and IL-8 in human peripheral blood mononuclear cells. We suggest that PR3 is a specific IL-32\u03b1 binding protein, independent of its enzymatic activity. However, limited cleavage of IL-32\u03b1 by PR3 enhances activities of the cytokine. Therefore, specific inhibition of PR3 activity to process IL-32 or neutralization of IL-32 by inactive PR3 or its fragments may reduce the consequences of IL-32 in immune regulated diseases.<\/jats:p>","DOI":"10.1073\/pnas.0511206103","type":"journal-article","created":{"date-parts":[[2006,2,17]],"date-time":"2006-02-17T20:34:03Z","timestamp":1140208443000},"page":"3316-3321","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":131,"title":["Proteinase 3 is an IL-32 binding protein"],"prefix":"10.1073","volume":"103","author":[{"given":"Daniela","family":"Novick","sequence":"first","affiliation":[{"name":"*Department of Molecular Genetics and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Menachem","family":"Rubinstein","sequence":"additional","affiliation":[{"name":"*Department of Molecular Genetics and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tania","family":"Azam","sequence":"additional","affiliation":[{"name":"Department of Medicine, University of Colorado Health Sciences Center, Denver, CO 80262"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aharon","family":"Rabinkov","sequence":"additional","affiliation":[{"name":"Unit of Biological Services, The Weizmann Institute of Science, Rehovot 76100, Israel, and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charles A.","family":"Dinarello","sequence":"additional","affiliation":[{"name":"Department of Medicine, University of Colorado Health Sciences Center, Denver, CO 80262"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Soo-Hyun","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Medicine, University of Colorado Health Sciences Center, Denver, CO 80262"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2006,2,17]]},"reference":[{"key":"e_1_3_3_1_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2004.12.003"},{"key":"e_1_3_3_2_2","doi-asserted-by":"crossref","first-page":"597","DOI":"10.4049\/jimmunol.148.2.597","volume":"148","author":"Dahl C. A.","year":"1992","unstructured":"C. A. Dahl, R. P. Schall, H. L. He, J. S. Cairns J. Immunol 148, 597\u2013603 (1992).","journal-title":"J. Immunol"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0508237102"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF01972395"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1097\/01.ASN.0000034911.03334.C3"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(94)00952-X"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2249.1996.d01-715.x"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.270.40.23688"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.96.11.6261"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.167.11.6568"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1006\/abbi.2001.2687"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00431-004-1464-3"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00059-004-2525-0"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199711203372106"},{"key":"e_1_3_3_15_2","first-page":"S64","volume":"21","author":"van Rossum A. P.","year":"2003","unstructured":"A. P. van Rossum, P. C. Limburg, C. G. Kallenberg Clin. Exp. Rheumatol 21, S64\u20138 (2003).","journal-title":"Clin. Exp. Rheumatol"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1523-1755.2004.00364.x"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1038\/nm968"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1681\/ASN.V1061224"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1097\/01.ASN.0000028642.26222.00"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI0213462"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1093\/rheumatology\/37.8.901"},{"key":"e_1_3_3_22_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1523-1755.2002.00619.x"},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.1097\/01.ASN.0000135058.46193.72"},{"key":"e_1_3_3_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(99)01098-4"},{"key":"e_1_3_3_25_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00408-003-1008-z"},{"key":"e_1_3_3_26_2","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.173.6.4179"},{"key":"e_1_3_3_27_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.170.4.1409"},{"key":"e_1_3_3_28_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)40049-5"},{"key":"e_1_3_3_29_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9673(01)93767-7"},{"key":"e_1_3_3_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(94)90154-6"},{"key":"e_1_3_3_31_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-7613(00)80013-8"},{"key":"e_1_3_3_32_2","doi-asserted-by":"publisher","DOI":"10.1210\/endo.138.8.5343"},{"key":"e_1_3_3_33_2","doi-asserted-by":"publisher","DOI":"10.1681\/ASN.V75694"},{"key":"e_1_3_3_34_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V93.3.849"},{"key":"e_1_3_3_35_2","first-page":"1617","volume":"149","author":"Yang J. J.","year":"1996","unstructured":"J. J. Yang, R. Kettritz, R. J. Falk, J. C. Jennette, M. L. Gaido Am. J. Pathol 149, 1617\u20131626 (1996).","journal-title":"Am. J. Pathol"},{"key":"e_1_3_3_36_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0002-9440(10)64000-X"},{"key":"e_1_3_3_37_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M400637200"},{"key":"e_1_3_3_38_2","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.165.11.6504"},{"key":"e_1_3_3_39_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI200316913"},{"key":"e_1_3_3_40_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0002-9440(10)64466-5"},{"key":"e_1_3_3_41_2","doi-asserted-by":"publisher","DOI":"10.1096\/fj.04-2469fje"},{"key":"e_1_3_3_42_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0409535102"},{"key":"e_1_3_3_43_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.82.4.1204"},{"key":"e_1_3_3_44_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V84.12.4242.bloodjournal84124242"},{"key":"e_1_3_3_45_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(88)90486-2"},{"key":"e_1_3_3_46_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-4274(94)90096-5"},{"key":"e_1_3_3_47_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI1379"},{"key":"e_1_3_3_48_2","doi-asserted-by":"publisher","DOI":"10.1152\/ajpregu.2001.281.4.R1264"},{"key":"e_1_3_3_49_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.231473998"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.0511206103","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,5,6]],"date-time":"2023-05-06T10:37:55Z","timestamp":1683369475000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.0511206103"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2006,2,17]]},"references-count":49,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2006,2,28]]}},"alternative-id":["10.1073\/pnas.0511206103"],"URL":"https:\/\/doi.org\/10.1073\/pnas.0511206103","relation":{"has-review":[{"id-type":"doi","id":"10.3410\/f.1009306.373825","asserted-by":"object"}]},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2006,2,17]]},"assertion":[{"value":"2006-02-17","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}