Separation
of Sulfated Nucleotides Using
Polyacrylamide Gel Electrophoresisl
Previous reports have indicated that nucleotide isolation and identification could be carried out using paper chromatography (l-3)) paper electrophoresis (4)) starch gel electrophoresis (5), and column chromatography (3,(6)(7)(8)(9)(10)(11). Separation techniques have also been established for thin-layer chromatography (11)(12)(13)(14)(15). A procedure for the rapid estimation of sulfated nucleotides has been developed in this laboratory using polyacrylamide gel electrophoresis for separation of 3'-phosphoadenosine 5'-phosphosulfate (PAPS) and adenosine 5'-phosphosulfate (APS) from inorganic sulfate in an in vitro reaction mixture. The presence of these compounds has previously been shown using a paper chromatographic systems (1,3). The use of polyacrylamide gel electrophoresis permits a more rapid and equally quantitative separation of the nucleotides produced from the in vitro synthesis of APS and PAPS from sulfate and ATP.
Methods and Materials. White Leghorn cockerels were raised from 'Approved by the Director of the West Virginia University Agricultural Experiment Station as Scientific Paper No. 1124. 'Communication No.