Capillary electroseparations of enkephalin-related peptides and protein kinase A peptide substrates.
The separations of enkephalin-related peptides and protein kinase A peptide substrates, with the common structural feature -Arg-Arg-X-Ser-Val-, were studied in micellar electrokinetic chromatography (MEKC) systems and compared with the capillary zone electrophoresis (CZE) mode. The influence of the magnitude and the direction of the electroosmotic flow on the selectivity was studied. Reversed electroosmosis was obtained by adding a hydrophobic amine, dimethyldodecylamine, to the background electrolyte; the amine forms cationic micelles with a low critical micelle concentration (0.3 mM). The neutral micellar agent, Brij 35, competes with the amine for adsorption sites on the capillary surface decreasing the reversed electroosmosis. In such a system, mixed cationic micelles are formed to which the peptides were not distributed at low pH, but an improved resolution was obtained due to the effects on electroosmosis. In systems containing the less hydrophobic amine dimethyloctylamine, in which probably no mixed micelles are formed, an improved separation of protein kinase A peptide substrates was obtained due to distribution to Brij 35 micelles. In separations of enkephalins, a high pH gave very low efficiencies due to surface-analyte interactions, and the best CZE separations were obtained at low pH. Changes in migration order were observed in the pH range 2-3, possibly due to differences in peptide pKa values or conformation changes of the peptides. The enkephalins were only to a small extent distributed to the Brij 35 micelles, but this improved the separation at pH 2 compared to the CZE mode.
Thorsteinsdóttir
M
M
Faculty of Pharmacy, Uppsala University Biomedical Centre, Uppsala, Sweden.
Beijersten
I
I
Westerlund
D
D
D000588:Amines / Q000737:chemistry; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002845:Chromatography / Q000379:methods; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004586:Electrophoresis / Q000379:methods*; D004745:Enkephalins / Q000032:analysis* / Q000737:chemistry; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis*; D000077423:Polidocanol; D011092:Polyethylene Glycols; D013379:Substrate Specificity
D000588:Amines; D000596:Amino Acids; D004745:Enkephalins; D010455:Peptides; D000077423:Polidocanol; D011092:Polyethylene Glycols; D017868:Cyclic AMP-Dependent Protein Kinases; C003935:dodecylamine