Assay and screening methods for bioactive substances based on cellular signaling pathways.
Assay and screening methods for bioactive substances based on cellular signaling pathways are presented. Examples include: (1) intracellular protein phosphorylation and protein-protein interaction, (1-i) a new assay method for evaluating chemical selectivity of agonists for insulin signaling pathways based on agonist-induced phosphorylation of a target peptide, (1-ii) an SPR-based screening method for agonist selectivity for insulin signaling pathways based on the binding of phosphotyrosine to its specific binding protein, (1-iii) a fluorescent indicator for tyrosine phosphorylation-based insulin signaling pathways, and (1-iv) split luciferase as an optical probe for detecting protein-protein interactions in mammalian cells based on protein splicing; (2) a screening method for antigen-specific IgE using mast cells based on intracellular calcium signaling; (3) a screening method for substrates of multidrug resistance-associated protein (MRP); and (4) fluorescent indicators for cyclic GMP based on cyclic GMP-dependent protein kinase Ialpha and green fluorescent proteins.
Umezawa
Yoshio
Y
Department of Chemistry, School of Science, The University of Tokyo, Hongo, Japan. umezawa@chem.s.u-tokyo.ac.jp
D018435:ATP Binding Cassette Transporter, Subfamily B / Q000737:chemistry / Q000378:metabolism; D000937:Antigen-Antibody Reactions; D001709:Biotechnology / Q000379:methods*; D006152:Cyclic GMP / Q000737:chemistry / Q000378:metabolism; D007073:Immunoglobulin E / Q000737:chemistry / Q000378:metabolism; D007328:Insulin / Q000737:chemistry / Q000378:metabolism; D008407:Mast Cells / Q000502:physiology; D010766:Phosphorylation; D011485:Protein Binding; D015398:Signal Transduction
D018435:ATP Binding Cassette Transporter, Subfamily B; D007328:Insulin; D007073:Immunoglobulin E; D006152:Cyclic GMP