Peptide mapping of the four subunits of the mouse DNA polymerase alpha-primase complex.
We report a simple, two-step method (phosphocellulose and immunoaffinity column chromatographies) for purification of the mouse DNA polymerase alpha-primase complex. The advantages of this method over other procedures are its simplicity and rapidity, with little loss by proteolysis. Sedimentation analysis in a glycerol density gradient of the immunoaffinity-purified fraction revealed that four polypeptides with molecular weights of 180,000, 68,000, 54,000 and 46,000 in the enzyme fraction form a physical complex. Peptide mapping by reversed phase-high performance liquid chromatography demonstrated unequivocally that these four polypeptides constituting the complex are different entities.
Takada-Takayama
R
R
Department of Physiological Chemistry, Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.
Tada
S
S
Hanaoka
F
F
Ui
M
M
D000818:Animals; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D019915:DNA Primase; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D010449:Peptide Mapping; D012316:RNA Nucleotidyltransferases / Q000302:isolation & purification*
D019915:DNA Primase; D012316:RNA Nucleotidyltransferases