Solid-phase sequence analysis of polypeptides eluted from polyacrylamide gels. An aid to interpretation of DNA sequences exemplified by the Escherichia coli unc operon and bacteriophage lambda.
An approach to sequencing proteins by the solid-phase method combined with isolation of proteins and polypeptides by gel electrophoresis is described. Mixtures of proteins or polypeptides resulting from digests are fractionated in the presence of dodecylsulphate in polyacrylamide gels. They are detected with Coomassie blue, eluted, selectively reacted with porous glass derivatives and sequenced in their amino-terminal regions with the aid of a new microsequencer. Alternatively they can be analysed or digested with enzymes and fingerprinted. It is a relatively rapid method of purifying proteins for sequence analysis which we have used to provide partial protein sequence data to complement DNA sequences. Nine genes, four from the unc operon of Escherichia coli encoding the alpha, beta, gamma and epsilon subunits of ATP synthase and five for capsid proteins of bacteriophage lambda, have been identified by this method.
D000251:Adenosine Triphosphatases / Q000235:genetics*; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000954:Antigens, Surface; D010582:Bacteriophage lambda / Q000235:genetics*; D001483:Base Sequence; D002213:Capsid / Q000235:genetics*; D004247:DNA; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D009876:Operon; D010446:Peptide Fragments; D010455:Peptides / Q000302:isolation & purification*; D010734:Phosphogluconate Dehydrogenase; D006180:Proton-Translocating ATPases; D014346:Trypanosoma brucei brucei / Q000276:immunology; D014764:Viral Proteins / Q000235:genetics*
D000596:Amino Acids; D000954:Antigens, Surface; D010446:Peptide Fragments; D010455:Peptides; D014764:Viral Proteins; D004247:DNA; D010734:Phosphogluconate Dehydrogenase; D000251:Adenosine Triphosphatases; D006180:Proton-Translocating ATPases