Identification of polypeptides on two-dimensional electrophoresis gels by amino acid composition.
We present a method that can, in principle, provide tentative identification of a substantial proportion of the polypeptides resolvable on a given two-dimensional electrophoresis gel. Computerized microdensitometry of autoradiograms from 20 gels labeled in turn with each of the 20 common amino acids provides the data for simultaneously measuring the amino acid composition of all polypeptides of interest on the gel. These compositions are then compared with computer data bases of known protein compositions. Similarity between a known and an unknown polypeptide with comparable molecular mass indicates a potential identification, which can then be confirmed with conventional techniques. We illustrate this technique by applying it to the identification of proteins in a transformed human cell line (HuT-14).
D000596:Amino Acids / Q000032:analysis*; D002460:Cell Line; D003201:Computers; D000242:Cyclic AMP / Q000494:pharmacology; D003720:Densitometry; D004586:Electrophoresis / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000032:analysis; D006801:Humans; D007525:Isoelectric Focusing; D009961:Orosomucoid / Q000032:analysis; D010455:Peptides / Q000032:analysis*; D011494:Protein Kinases / Q000032:analysis; D011498:Protein Precursors / Q000032:analysis; D006180:Proton-Translocating ATPases / Q000032:analysis; D014335:Tropomyosin / Q000032:analysis
D000596:Amino Acids; D009961:Orosomucoid; D010455:Peptides; D011498:Protein Precursors; D014335:Tropomyosin; D000242:Cyclic AMP; D011494:Protein Kinases; D006180:Proton-Translocating ATPases