Ion-exchange chromatography of ionic detergent-solubilized proteins: application to purification of rat parotid gland phosphoproteins including ribosomal protein S6.
A method is described for separation of ionic detergent-solubilized proteins by ion-exchange chromatography. This method has been developed for purification of two phosphoproteins (Mr 19,000 and 30,000) from 32Pi-prelabeled, isoproterenol-stimulated rat parotid tissue and is based on the observation that, in the presence of urea and Nonidet-P40, ionic detergent-solubilized proteins can be adsorbed by ion exchangers according to their own charge. After adsorption, proteins were eluted with a stepwise gradient of NaCl in a urea-containing buffer. By the procedure described, the 30 kDa phosphoprotein was freed from other 32P-labeled substances; and it was identified as ribosomal protein S6 that was phosphorylated at some serine residues. The method is generally applicable and especially suited for preliminary purification of hydrophobic proteins subjected to analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
Department of Biochemistry, School of Dentistry, Aichi-Gakuin University, Nagoya, Japan.
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Kanamori
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Hayakawa
D000818:Animals; D000077286:Cetrimonium; D002593:Cetrimonium Compounds; D002848:Chromatography, DEAE-Cellulose / Q000379:methods*; D008970:Molecular Weight; D010306:Parotid Gland / Q000032:analysis*; D010750:Phosphoproteins / Q000302:isolation & purification*; D051381:Rats; D038601:Ribosomal Protein S6; D012269:Ribosomal Proteins / Q000302:isolation & purification; D012967:Sodium Dodecyl Sulfate; D012995:Solubility
D002593:Cetrimonium Compounds; D010750:Phosphoproteins; D038601:Ribosomal Protein S6; D012269:Ribosomal Proteins; D012967:Sodium Dodecyl Sulfate; D000077286:Cetrimonium