Autoimmune antigen Ku is enriched on oligonucleotide columns distinct from those containing the octamer binding protein DNA consensus sequence.
During purification of the AP1 complex from the T cell line MLA144 we enriched for a complex which bound to an oligonucleotide column containing the AP1 DNA consensus sequence and co-eluted with a fraction required for AP1 binding activity. This complex although co-eluting with AP1 binding activity had previously been determined to be non-specific in its DNA binding properties. Further investigation determined that the complex was a heterodimer of 85 and 70 kDa which was antigenically related to the autoimmune antigen Ku. It is important to be aware of the abundance and avidity of the Ku complex to bind oligonucleotide columns when purifying sequence specific binding proteins.
Quinn
J P
JP
MRC Brain Metabolism Unit, Royal Edinburgh Hospital, Scotland.
Farina
A R
AR
D000818:Animals; D034961:Antigens, Nuclear; D001483:Base Sequence; D015153:Blotting, Western; D002846:Chromatography, Affinity; D016384:Consensus Sequence; D004247:DNA / Q000737:chemistry; D004265:DNA Helicases; D004268:DNA-Binding Proteins / Q000737:chemistry*; D000072200:Ku Autoantigen; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry*; D009841:Oligonucleotides; D014407:Tumor Cells, Cultured
D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D009841:Oligonucleotides; D004247:DNA; D004265:DNA Helicases; C106893:XRCC5 protein, human; C000606509:Xrcc6 protein, human; D000072200:Ku Autoantigen