Electromigration behavior of poly-(L-glutamate) conformers in concentrated polyacrylamide gels.
During electrophoretic separation of anionic polyamino acids, resolution according to the number of peptide units can be achieved in capillaries filled with hydrophilic gels. While polyaspartate preparations yield single peaks for the individual oligomers at pH above 8.0, polyglutamates exhibit an anomalous behavior of peak splitting, which is attributed here to the separation of the oligopeptide conformers. An Asp-Glu (1:1) copolymer yields single peaks under similar conditions. At pH near 4.5, where polyglutamate is expected to exist in its alpha-helix form, peak splitting disappears. Upon heating to 95 degrees C for at least 120 h (procedure described to transform the alpha-helix into a beta form), peak splitting disappeared, but could be reestablished after cooling for several days. When a highly charged cation spermine was added to the operational electrolyte, triple peaks appeared in the electropherogram due to the ion-pair formation. The largest peak in every triplet has been tentatively assigned to the alpha-helix form. The electrophoretic results described have been largely supported by CD spectra.
Dolník
V
V
Department of Chemistry, Indiana University, Bloomington.
Novotny
M
M
Chmelík
J
J
D000180:Acrylic Resins / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D011099:Polyglutamic Acid / Q000737:chemistry*; D011487:Protein Conformation
D000180:Acrylic Resins; C016680:polyacrylamide gels; D011099:Polyglutamic Acid