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When incubated at low concentrations (&lt; 1 n<jats:sc>M<\/jats:sc>) <jats:sup>125<\/jats:sup>I\u2010labeled TNF\u2010\u03b1 and unlabeled TNF\u2010\u03b1 rapidly multimerized or dissociated into monomers and bioactivity decreased. Sodium dodecyl sulfate (SDS)\u2010PAGE analysis of cross\u2010linked <jats:sup>125<\/jats:sup>I\u2010labeled TNF\u2010\u03b1 demonstrated bands of multi\u2010 and trimeric TNF\u2010\u03b1 in addition to dominating bands of dimers and monomers. Tri\u2010, di\u2010 and monomeric TNF\u2010\u03b1 were recovered from SDS\u2010PAGE gels and allowed to renature. Of the original receptor\u2010binding activity, 10%\u201315% was obtained with cross\u2010linked TNF\u2010\u03b1 dimers, whereas none was recovered from preparations of trimeric TNF\u2010\u03b1. Multimeric and monomeric TNF\u2010\u03b1 exhibited little or no binding activity, and cell\u2010bound, cross\u2010linked TNF\u2010\u03b1 which was dissociated from cellular binding sites was mainly dimeric. <jats:sup>125<\/jats:sup>I\u2010labeled TNF\u2010\u03b1 bound to lymphokine\u2010activated killer (LAK) cells and binding kinetics were much similar (K<jats:sub>d<\/jats:sub> approximately 100 p<jats:sc>M<\/jats:sc>) to those reported in other normal cell types. The number of receptors per LAK cell was approximately 4 \u00d7 10<jats:sup>3<\/jats:sup>. Cross\u2010linking of TNF\u2010\u03b1 to binding sites in U\u2010937 and LAK cells yielded a receptor\u2010ligand complex of about 80\/90 kDa. At 37 \u00b0C, <jats:sup>125<\/jats:sup>I\u2010labeled TNF\u2010\u03b1 was rapidly internalized and degraded in L\u2010929, U\u2010937 and LAK cells. Degradation of ligand and recycling of receptors were blocked in the presence of methylamine. Methylamine significantly inhibited TNF\u2010\u03b1\u2010mediated cytolysis of L\u2010929 cells and caused a quantitatively corresponding reduction in cellular TNF\u2010\u03b1 uptake, indicating that L\u2010929 lysis was mediated by receptors.<\/jats:p>","DOI":"10.1002\/eji.1830191020","type":"journal-article","created":{"date-parts":[[2007,3,1]],"date-time":"2007-03-01T08:33:43Z","timestamp":1172738023000},"page":"1887-1894","source":"Crossref","is-referenced-by-count":27,"title":["Bioactive human recombinant tumor necrosis factor\u2010\u03b1: an unstable dimer?"],"prefix":"10.1002","volume":"19","author":[{"given":"Claus Munck","family":"Petersen","sequence":"first","affiliation":[]},{"given":"Anders","family":"Nykj\u00e6r","sequence":"additional","affiliation":[]},{"given":"Brian S.","family":"Christiansen","sequence":"additional","affiliation":[]},{"given":"Lene","family":"Heickendorff","sequence":"additional","affiliation":[]},{"given":"S\u00f8Ren C.","family":"Mogensen","sequence":"additional","affiliation":[]},{"given":"Bjarne","family":"M\u00f8ller","sequence":"additional","affiliation":[]}],"member":"311","published-online":{"date-parts":[[2005,11,22]]},"reference":[{"key":"e_1_2_1_2_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.72.9.3666"},{"key":"e_1_2_1_3_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.0954-6820.1986.tb02785.x"},{"key":"e_1_2_1_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/0167-5699(85)90144-6"},{"key":"e_1_2_1_5_2","first-page":"235","volume":"2","author":"Ruff M. 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