Cayman Chemical Company Incorporated

University of Michigan

Jeffrey Keith Johnson

Gregory William Endres

Stephen Douglas Barrett

Fred Lawrence Ciske

Raymond C Trievel

Levi Lynn Blazer

Daniel Austin Bochar

Margaret Lynn Collins

Nisha Palackal

Karie L McGowan

Gregory Scott Keyes

Kirk W Maxey

Ranjinder Singh Sidhu

Assay methods may generally comprise forming homogeneous assay mixtures comprising target SAM-utilizing protein, fluorescent detection analyte, and test compound, incubating, and measuring FP or TR-FRET signal emitted in order to determine a measure of test compound-SAM -utilizing protein binding. Assay mixtures comprise a SAM-utilizing protein, and a fluorescent detection analyte that binds with the SAM-utilizing protein in the absence of test compound. Assay mixtures may further comprise a test compound. Assay mixture embodiments may generate FP or TR-FRET signal properties that are a function of the inherent binding interactions of both the test compound and the detection analyte with the SAM-utilizing protein. Fluorescent detection analytes comprise a fluorophore moiety, a covalent linker moiety, and a SAM-utilizing protein ligand moiety and could be utilized in FP or TR-FRET assays to measure test compound binding.

Fluorescent molecular probes for use in assays that measure test compound competitive binding with SAM-utilizing proteins

An exemplary embodiment may be directed to a fluorescence polarization assay that screens compounds or agents for their affinity to hematopoietic prostaglandin D synthase (H-PGDS) based on their ability to displace a fluorophore-containing detection analyte bound to an enzyme comprising the primary amino acid sequence of H-PGDS. Another exemplary embodiment utilizes an enzyme having a maltose binding protein amino-acid sequence fused with an N-terminus of the enzyme.

Method for assaying compounds or agents for ability to displace potent ligands of hematopoietic prostaglandin D synthase

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Ann Arbor, MI, United States of America

Jeffrey Keith Johnson