title: Covalent cross-links in oxygen free radical altered human immunoglobulin G.
abstract: The damaging effect of an oxygen free radical generating system, i.e. ultraviolet irradiation, on human immunoglobulin G (IgG) was studied. The free radical altered IgG was analysed by a high performance liquid chromatograph equipped with a TSK G 3000 SW-column. Gel filtration of 120 min UV-irradiated IgG resulted in three clearly distinguished peaks corresponding to polymer IgG (MW greater than 500 kD), dimer IgG (MW 300 kD) and monomer IgG (MW 150 kD). Analysis of oxygen free radical altered and aggregated IgG by SDS-PAGE and subsequent silver-staining revealed inter- and intra-molecular reduction (by beta-mercaptoethanol)-resistant cross-links between IgG-molecules were formed. Comparison of amino acid analyses of native IgG with oxygen free radical aggregated polymer IgG showed significant reductions in tyrosine- (7.0%) and histidine- (6.5%) content. These findings suggest that tyrosine and histidine are involved in covalent cross-linking between IgG-molecules caused by oxygen free radicals. These alterations on IgG induced by free radical-activity might render it antigenic, and could initiate the production of rheumatoid factors (RF).
authors.affiliation: Department of Biochemistry I, Medical Faculty, Erasmus University, Rotterdam, The Netherlands.
authors.forename: H A
authors.initials: HA
authors.lastname: Kleinveld
authors.forename: C E
authors.initials: CE
authors.lastname: Hack
authors.forename: A J
authors.initials: AJ
authors.lastname: Swaak
authors.forename: W L
authors.initials: WL
authors.lastname: van Noort
authors.forename: H G
authors.initials: HG
authors.lastname: van Eijk
authors.forename: J F
authors.initials: JF
authors.lastname: Koster
mesh_terms: D000596:Amino Acids / Q000032:analysis; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D005609:Free Radicals; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D010100:Oxygen / Q000494:pharmacology*; D014466:Ultraviolet Rays
chemical_list: D000596:Amino Acids; D003432:Cross-Linking Reagents; D005609:Free Radicals; D007074:Immunoglobulin G; D010100:Oxygen