University of California

Sequenom, Inc.

Susan S Taylor

Roger Y Tsien

Charles Cantor

Gary Olson

Christopher Self

Andreas Braun

Charles Michael Cook

Stephen R Adams

Ying Ji

Stefan M Kammerer

Sanjay A Saldanha

Lora Burns-Hamuro

Methods for screening compounds for their activity as a protein kinase A modulators are provided. The methods are based on fluorescence polarization of peptide probes to identify drug candidates that act by activating or inhibiting the catalytic function of PKA. In certain embodiments, the methods are adapted for high throughput screening.

Screening assay to identify modulators of protein kinase A

A-kinase anchor protein (AKAPs) muteins, peptides thereof, and nucleic acids encoding the peptides are provided herein. Also provided are transgenic animals, cells comprising transgenes and various methods employing such peptides.

Kinase anchor protein muteins, peptides thereof and related documents

The invention provides labeled proteins suitable for determining the presence of cAMP, other second messengers and organic molecules. The proteins are separately labeled with fluorochromes which, when in close spatial proximity, preferably, less than about 6 nm, interact through the transfer of energy from one fluorochrome to the other. A composition of matter, (S.sub.1.A).sub.n1 (S.sub.2.D).sub.n2 is provided wherein S.sub.1 and S.sub.2 are two proteins which are associated in one state and substantially disassociated in another, the equilibrium between which is controlled by the free concentration of an analyte, and A and D are fluorochromes, the emission wavelength of fluorochrome D overlapping the excitation wavelength of fluorochrome A and the distance between A and D being in sufficiently close proximity to allow the radiationless transfer of energy between the fluorochromes. A and D can be selected from a variety of acceptor, donor pairs such as fluorescein and tetramethylrhodamine and derivatives thereof. The concentration of analytes such as cAMP in a sample can be determined by contacting the sample with (S.sub.1.A).sub.n1 (S.sub.2.D).sub.n2 providing energy near the excitation wavelength of D and measuring the fluorescence of A or D, the concentration of cAMP and other said analytes being determined by the ratio of emission of D to the emission of A, previously calibrated with reference solutions of known analyte concentration.

Detection of analytes using fluorescent energy transfer

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Del Mar, CA, United States of America

Susan S Taylor