Gamble T N, Ramachandran C, Bateman K P
Merck Frosst Canada & Co., Kirkland, Quebec, Canada.
Anal Chem. 1999 Aug 15;71(16):3469-76. doi: 10.1021/ac990276t.
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.
利用带有紫外检测的毛细管区带电泳（CZE - UV）开发了快速分离磷酸肽异构体（具有相同序列但磷酸基团位于不同残基上的肽）的方法。未涂层、阳离子和中性毛细管与酸性和碱性肽一起使用。这些方法能够检测几种蛋白激酶（丝裂原活化蛋白激酶、蛋白激酶A、谷胱甘肽 - S - 转移酶 - 酪氨酸激酶）和磷酸酶（酸性、碱性和蛋白酪氨酸磷酸酶），并确定磷酸化和去磷酸化位点。非磷酸化或磷酸化肽与激酶或磷酸酶的孵育直接在仪器的自动进样器中进行，并使用CZE - UV在数小时内进行监测。
