{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,19]],"date-time":"2025-09-19T10:43:50Z","timestamp":1758278630244},"reference-count":32,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2005,12,1]],"date-time":"2005-12-01T00:00:00Z","timestamp":1133395200000},"content-version":"vor","delay-in-days":6209,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Eur J Immunol"],"published-print":{"date-parts":[[1988,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The human type I interferon (IFN) receptor was characterized by ligand blotting. In this method, plasmalemma proteins or detergent\u2010lysed whole\u2010cell extracts from human Burkitt lymphoma cell lines were separated on polyacrylamide gels and subsequently transferred onto nitrocellulose sheets. Probing the blots with 3 \u00d7 10<jats:sup>\u221210<\/jats:sup> M <jats:sup>125<\/jats:sup>I\u2010labeled recombinant IFN\u2010\u03b1A (<jats:sup>125<\/jats:sup>I\u2010rIFN\u2010\u03b1A) revealed an IFN\u2010\u03b1\u2010binding protein with an apparent molecular mass of 95 kDa (p95). Performing the electrophoretic run under reducing conditions completely abrogated the signal on the blot, indicating that the type I IFN receptor contains a'disulfide bond essential for IFN binding. Optimal binding of <jats:sup>125<\/jats:sup>I\u2010rIFN\u2010\u03b1A occurred at pH 9. The specificity of the binding reaction was established by simultaneously adding an excess of unlabeled IFN during incubation of the blots with <jats:sup>125<\/jats:sup>I\u2010rIFN\u2010\u03b1A. The addition of either unlabeled IFN\u2010\u03b1 or IFN\u2010\u03b2, but not IFN\u2010\u03b3, abolished the binding of <jats:sup>125<\/jats:sup>I\u2010rIFN\u2010\u03b1A to p95. <jats:sup>125<\/jats:sup>I\u2010rIFN\u2010\u03b3 at 1.25 \u00d7 10<jats:sup>\u221211<\/jats:sup> M bound to two proteins distinct from p95 with apparent molecular mass of 92 and 87 kDa, respectively. Saturability of <jats:sup>125<\/jats:sup>I\u2010rIFN\u2010\u03b1A binding was demonstrated by probing a constant amount of membrane proteins with increasing amounts of <jats:sup>125<\/jats:sup>I\u2010rIFN\u2010\u03b1A. Scatchard analysis of the binding data yielded an apparent K<jats:sub>d<\/jats:sub> of 5.4 \u00d7 10<jats:sup>\u221210<\/jats:sup> M for the immobilized type I IFN receptor. The expression of p95 on IFN\u2010\u03b1\u2010resistant and \u2010sensitive cells was indistinguishable. We conclude that p95 is the IFN\u2010\u03b1\/\u03b2 receptor and that two proteins (p92 and p87) can specifically bind IFN\u2010\u03b3. These results indicate that ligand blotting is a versatile method for characterization of unmodified IFN receptors and IFN\u2010receptor interaction and could also provide a new investigational approach for other cytokine receptor systems.<\/jats:p>","DOI":"10.1002\/eji.1830181221","type":"journal-article","created":{"date-parts":[[2007,3,1]],"date-time":"2007-03-01T01:04:15Z","timestamp":1172711055000},"page":"2009-2014","source":"Crossref","is-referenced-by-count":19,"title":["Characterization of the human type I interferon receptor by ligand blotting"],"prefix":"10.1002","volume":"18","author":[{"given":"Michael","family":"Schwabe","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gerald L.","family":"Princler","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Connie R.","family":"Faltynek","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2005,12]]},"reference":[{"key":"e_1_2_1_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/0163-7258(84)90037-8"},{"key":"e_1_2_1_3_2","first-page":"81","volume":"1","author":"Faltynek C. R.","year":"1984","journal-title":"Microbiol. Sci."},{"key":"e_1_2_1_4_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-065X.1987.tb00515.x"},{"key":"e_1_2_1_5_2","first-page":"61","volume":"7","author":"Celada A.","year":"1988","journal-title":"Lymphokine Res."},{"key":"e_1_2_1_6_2","doi-asserted-by":"publisher","DOI":"10.1038\/294768a0"},{"key":"e_1_2_1_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)33444-6"},{"key":"e_1_2_1_8_2","first-page":"13","volume":"257","author":"Branca A. A.","year":"1982","journal-title":"J. Biol. Chem."},{"key":"e_1_2_1_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/0042-6822(83)90127-7"},{"key":"e_1_2_1_10_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.85.13.4837"},{"key":"e_1_2_1_11_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.165.4.988"},{"key":"e_1_2_1_12_2","doi-asserted-by":"crossref","first-page":"8483","DOI":"10.1016\/S0021-9258(18)47439-X","volume":"262","author":"Novick D.","year":"1987","journal-title":"J. Bioi. Chem."},{"key":"e_1_2_1_13_2","doi-asserted-by":"crossref","first-page":"4231","DOI":"10.4049\/jimmunol.140.12.4231","volume":"140","author":"Sheehan K. C. F.","year":"1988","journal-title":"J. Immunol."},{"key":"e_1_2_1_14_2","doi-asserted-by":"publisher","DOI":"10.1089\/jir.1987.7.683"},{"key":"e_1_2_1_15_2","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/S0021-9258(19)45311-8","volume":"257","author":"Joshi A. R.","year":"1982","journal-title":"J. Biol. Chem."},{"key":"e_1_2_1_16_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1986.tb09655.x"},{"key":"e_1_2_1_17_2","doi-asserted-by":"crossref","first-page":"8017","DOI":"10.1016\/S0021-9258(19)57505-6","volume":"261","author":"Zhang Z.","year":"1986","journal-title":"J. Biol. Chem."},{"key":"e_1_2_1_18_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1988.tb13840.x"},{"key":"e_1_2_1_19_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1988.tb13839.x"},{"key":"e_1_2_1_20_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.80.11.3269"},{"key":"e_1_2_1_21_2","doi-asserted-by":"publisher","DOI":"10.1002\/jcp.1041230325"},{"key":"e_1_2_1_22_2","doi-asserted-by":"crossref","first-page":"4134","DOI":"10.4049\/jimmunol.136.11.4134","volume":"136","author":"Faltynek C. R.","year":"1986","journal-title":"J. Immunol."},{"key":"e_1_2_1_23_2","doi-asserted-by":"publisher","DOI":"10.1042\/bj1680187"},{"key":"e_1_2_1_24_2","doi-asserted-by":"publisher","DOI":"10.1038\/227680a0"},{"key":"e_1_2_1_25_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.76.9.4350"},{"key":"e_1_2_1_26_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI112228"},{"key":"e_1_2_1_27_2","first-page":"5339","volume":"46","author":"Uecer U.","year":"1986","journal-title":"Cancer Res."},{"key":"e_1_2_1_28_2","doi-asserted-by":"crossref","first-page":"1191","DOI":"10.1016\/S0021-9258(20)71226-3","volume":"260","author":"Littman S. J.","year":"1985","journal-title":"J. Biol. Chem."},{"key":"e_1_2_1_29_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.80.16.4973"},{"key":"e_1_2_1_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)32666-8"},{"key":"e_1_2_1_31_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)47584-9"},{"key":"e_1_2_1_32_2","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(87)90142-4"},{"key":"e_1_2_1_33_2","doi-asserted-by":"publisher","DOI":"10.1016\/0303-7207(87)90061-X"}],"container-title":["European Journal of Immunology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Feji.1830181221","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/eji.1830181221","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,22]],"date-time":"2023-10-22T01:17:00Z","timestamp":1697937420000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/eji.1830181221"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1988,12]]},"references-count":32,"journal-issue":{"issue":"12","published-print":{"date-parts":[[1988,12]]}},"alternative-id":["10.1002\/eji.1830181221"],"URL":"https:\/\/doi.org\/10.1002\/eji.1830181221","archive":["Portico"],"relation":{},"ISSN":["0014-2980","1521-4141"],"issn-type":[{"value":"0014-2980","type":"print"},{"value":"1521-4141","type":"electronic"}],"subject":[],"published":{"date-parts":[[1988,12]]}}}