Phosphopeptide Isomer Separation Using Capillary Zone Electrophoresis for the Study of Protein Kinases and Phosphatases
- 16 July 1999
- journal article
- research article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 71 (16), 3469-3476
- https://doi.org/10.1021/ac990276t
Abstract
Methods for the rapid separation of phosphopeptide isomers (peptides with the same sequence but with phosphates on different residues) were developed using capillary zone electrophoresis with ultraviolet (CZE-UV) detection. Uncoated, cationic and neutral capillaries were used with both acidic and basic peptides. These methods enabled the assay of several protein kinases (mitogen activated protein kinase, protein kinase A, GST-tyrosine kinase) and phosphatases (acid, alkaline, and protein tyrosine phosphatase) and the determination of the sites of phosphorylation and dephosphorylation. Incubations of nonphosphorylated or phosphorylated peptide with kinases or phosphatases took place directly in the instrument's autosampler and were monitored over several hours using CZE-UV.Keywords
This publication has 20 references indexed in Scilit:
- Phosphopeptide Analysis by Matrix-Assisted Laser Desorption Time-of-Flight Mass SpectrometryAnalytical Chemistry, 1996
- Phosphorylation and Activation of a cAMP-specific Phosphodiesterase by the cAMP-dependent Protein KinaseJournal of Biological Chemistry, 1996
- High‐sensitivity determination of tyrosine‐phosphorylated peptides by on‐line enzyme reactor and electrospray ionization mass spectrometryProtein Science, 1995
- Direct Determination of the Sequence Recognition Requirements of the SH2 Domains of SH-PTP2Biochemistry, 1995
- Protein kinases and phosphatases: The Yin and Yang of protein phosphorylation and signalingCell, 1995
- Capillary electrophoretic resolution of phosphorylated peptide isomers using micellar solutions and coated capillariesElectrophoresis, 1995
- Cdc2 regulatory factorsCurrent Opinion in Cell Biology, 1994
- Monitoring protein kinase and phosphatase reactions with matrix-assisted laser desorption/ionization mass spectrometry and capillary zone electrophoresis: Comparison of the detection efficiency of peptide-phosphopeptide mixturesJournal of Mass Spectrometry, 1994
- Phosphopeptide mapping and phosphoamino acid analysis by electrophoresis and chromatography on thin‐layer cellulose platesElectrophoresis, 1994
- Capillary electrophoresis: pioneering new approaches for biomolecular analysisTrends in Biochemical Sciences, 1993