Capillary zone electrophoresis and micellar electrokinetic chromatography, with taurodeoxycholate as micellar agent, of protein kinase A peptide substrates.
The separation of protein kinase A peptide substrates with the general formula -X-Arg-Arg-Ala-Ser-Y-, where X and Y may be the same or different amino acids, was studied by capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC). Taurodeoxycholate (TDC) was used as the micellar agent. CZE was effective in separating a peptide series differing in the number of amino acids, but not for a series with a difference in the terminating amino acid. For the latter series, MEKC generally gave a higher selectivity, but some of the peptide pairs were more easily separated by CZE, demonstrating the complementary character of the two techniques. The efficiency of the MEKC system was typically < 50% of that of CZE, but its higher selectivity generally outbalanced the lower efficiency regarding resolution. The distribution of the peptides to the micelles was studied by determination of retention factors. Electrostatic and hydrophobic forces were found to be determining factors in the distribution; the most highly charged basic peptides were most heavily distributed, and for peptides with the same charge those containing more hydrophobic amino acids were more strongly distributed. The contribution of some structural features to the distribution degree was also determined.
Analytical Pharmaceutical Chemistry, Faculty of Pharmacy, Uppsala University, Sweden.
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I
Beijersten
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D
Westerlund
D000595:Amino Acid Sequence; D002845:Chromatography / Q000379:methods*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry*; D004563:Electrochemistry; D019075:Electrophoresis, Capillary; D008823:Micelles; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010446:Peptide Fragments / Q000032:analysis*; D013379:Substrate Specificity; D013501:Surface-Active Agents; D013657:Taurodeoxycholic Acid
D008823:Micelles; D010446:Peptide Fragments; D013501:Surface-Active Agents; D013657:Taurodeoxycholic Acid; D017868:Cyclic AMP-Dependent Protein Kinases