title: Assays using a NIMS chip: loosely bound but highly selective.
abstract: A fluorous-affinity nanostructure initiator mass spectrometry (NIMS) chip has been developed as an enzyme activity assay. As a proof-of-principle, an assay for detecting cysteine-containing peptide phosphorylation by protein kinase A (PKA) was designed using NIMS technology. The efficacy for an enzyme inhibition assay was characterized by deriving an IC50 value from the ratio of the substrate-to-product mass spectral signal intensities, using known inhibitors of PKA and Abl kinase activity. Lastly, the potential use of a NIMS chip as a multiple screening enzyme inhibition platform was explored.
authors.affiliation: Department of Chemistry, University of Victoria, Stn CSC, Victoria, BC, Canada.
authors.forename: J
authors.initials: J
authors.lastname: Li
authors.forename: R H
authors.initials: RH
authors.lastname: Lipson
mesh_terms: D057075:Enzyme Assays / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D049329:Nanostructures / Q000737:chemistry*; D010446:Peptide Fragments / Q000032:analysis* / Q000378:metabolism; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding / Q000502:physiology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
chemical_list: D010446:Peptide Fragments