title: Activity measurement of protein kinase and protein phosphatase by microchip phosphate-affinity electrophoresis.
abstract: We previously proposed microchip-based phosphate-affinity electrophoresis (μPAE) and demonstrated its application to activity measurement of a tyrosine kinase, c-Src. In this study, we extended the μPAE application to a serine/threonine kinase, protein kinase A (PKA), and to a tyrosine phosphatase, leukocyte antigen-related protein tyrosine phosphatase (LAR PTPase). For standard peptide samples, we obtained linear calibration plots, and the limits of detection were 1.2% (PKA) and 1.5% (LAR PTPase) product peptides in the total peptides. The μPAE was also proven to be effective for unpurified enzyme reaction products.
authors.affiliation: Bioengineering Laboratory, RIKEN, Wako, Saitama 351-0198, Japan.
authors.forename: Aishan
authors.initials: A
authors.lastname: Han
authors.forename: Kazuo
authors.initials: K
authors.lastname: Hosokawa
authors.forename: Mizuo
authors.initials: M
authors.lastname: Maeda
mesh_terms: D004586:Electrophoresis / Q000379:methods*; D046209:Electrophoresis, Microchip / Q000379:methods*; D010710:Phosphates / Q000378:metabolism*; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*
chemical_list: D010710:Phosphates; D011494:Protein Kinases; D010749:Phosphoprotein Phosphatases