Complex protein binding within the mouse immunoglobulin heavy-chain enhancer.
We have begun to purify and characterize several proteins which bind to the mouse immunoglobulin heavy-chain enhancer to understand the molecular interactions important for enhancer activity. Three proteins which bind to different sites on the immunoglobulin heavy-chain enhancer have been chromatographically separated and partially purified. One protein binds a site which has not been reported previously and does not bind to other reported protein-binding sites on the immunoglobulin heavy-chain enhancer. Binding-site boundaries for the three partially purified proteins have been precisely mapped by methylation interference, DNase I footprinting, and orthophenanthroline/copper chemical nuclease footprinting. We have also characterized these three proteins with respect to dissociation rate constants.
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002467:Cell Nucleus / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D003300:Copper; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D003850:Deoxyribonuclease I; D004591:Electrophoresis, Polyacrylamide Gel; D004742:Enhancer Elements, Genetic; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007202:Indicators and Reagents; D008745:Methylation; D051379:Mice; D008969:Molecular Sequence Data; D010618:Phenanthrolines; D010954:Plasmacytoma / Q000032:analysis; D011485:Protein Binding
D004268:DNA-Binding Proteins; D007143:Immunoglobulin Heavy Chains; D007202:Indicators and Reagents; D010618:Phenanthrolines; D003300:Copper; D004247:DNA; D003850:Deoxyribonuclease I; C025205:1,10-phenanthroline