The disulphide bridges of a mouse immunoglobulin G1 protein.
[(35)S]Cystine-labelled immunoglobulin MOPC21 (IgG1) was prepared from myeloma cells in tissue culture. Carrier myeloma protein was added and the protein was digested with pepsin. The digest was fractionated on Sephadex G-50 into two fractions, further digested with trypsin and again fractionated on Sephadex. Disulphide-bridge peptides were purified by electrophoresis and chromatography and identified by radioautography. A peptide of 96 residues was isolated, which contains both the heavy-light interchain disulphide bridge and all the inter-heavy-chain disulphide bridges. Other peptides were isolated, accounting for all the intrachain disulphide bridges (which could be placed by homology with proteins of other species), except for the variable section of the light chain. Sequences describing this missing disulphide bridge were obtained from totally reduced and alkylated light chains. Peptides related to the interchain disulphide-bridge peptide were isolated from partially reduced and alkylated myeloma protein and from totally reduced heavy chain. The interchain disulphide-bridge peptide was placed at the C-terminal position of the F(ab')(2) fragment, prepared by digestion of the protein with pepsin at pH4.0. Sequences from the heavy-chain intrachain disulphide bridges of MOPC 21 immunoglobulin are compared with homologous sequences from mouse myeloma proteins of other subclasses and proteins of other species.
D000595:Amino Acid Sequence; D000818:Animals; D001345:Autoradiography; D002247:Carbon Isotopes; D002848:Chromatography, DEAE-Cellulose; D046508:Culture Techniques; D003553:Cystine / Q000378:metabolism; D004220:Disulfides / Q000032:analysis; D004590:Electrophoresis, Paper; D007122:Immunoelectrophoresis; D007074:Immunoglobulin G / Q000032:analysis; D007136:Immunoglobulins / Q000032:analysis*; D007461:Iodoacetates / Q000378:metabolism; D051379:Mice; D009194:Myeloma Proteins / Q000032:analysis; D010084:Oxidation-Reduction; D013440:Sulfides / Q000032:analysis*; D013460:Sulfur Isotopes; D014357:Trypsin
D002247:Carbon Isotopes; D004220:Disulfides; D007074:Immunoglobulin G; D007136:Immunoglobulins; D007461:Iodoacetates; D009194:Myeloma Proteins; D013440:Sulfides; D013460:Sulfur Isotopes; D003553:Cystine; D014357:Trypsin