Bellbrook Labs, LLC

Panvera LLC

Robert G Lowery

Karen Kleman-Leyer

Matt Staeben

Thane Westermeyer

Thomas J Burke

David V Thompson

Peter M Shaw

Randall E Bolger

William J Checovich

John Majer

Olga Trubetskoy

Thomas K Zielinski

Kerry M Ervin

The present invention relates to novel engineered Ga proteins and assay methods of using such proteins to advance drug discovery. Engineered Ga proteins described by the invention contain alterations of at least one and preferably two or more amino acid residues that are highly conserved among all four subfamilies of Ga proteins. A preferred engineered protein disclosed here is a double mutant, G&#x3b1;&#x3c0; R178M A326S. This specific combination of mutations yields an unexpectedly amplified effect on Ga function both in terms of GTPase activity (GTP hydrolysis) and GDP dissociation. This synergistic effect may have a profound influence on the way GPCR signaling pathways are examined for the development of new pharmacotherapies, particularly in the field of central nervous system disorders such as Parkinson's disease.

Genetically engineered G-alpha proteins and uses thereof

An isolated antibody produced against a donor product of an enzymatically catalyzed group transfer reaction is disclosed. In one aspect, the antibody has the ability to preferentially recognize the donor-product guanosine diphosphate (GDP), in the presence of the donor molecule guanosine triphosphate (GTP). Immunuoassays using compositions including such antibodies and kits for performing the immunoassays are also disclosed.

Assay method for group transfer reactions

The present invention relates to methods for detecting, quantifying and high throughput screening of donor-products and the catalytic activities generating the donor-products in group-transfer reactions. The invention further provides immunoassays, antibodies and kits that may be used to practice the methods of the invention.

Antibody that binds uridine diphosphate

The present invention relates to methods for detecting, quantifying and high throughput screening of donor-products and the catalytic activities generating the donor-products in group-transfer reactions catalyzed by adenosine triphosphatase (ATPase) or guanine triphosphatase (GTPase). The invention further provides immunoassays, antibodies and kits that may be used to practice the methods of the invention.

Assay methods for group transfer reactions

A fluorescence polarization process used to identify activity of conjugative enzymes involved in xenobiotic transformations, such as glucuronosyltransferases is provided.

Process for glucuronidation screening

The system comprises mixing a fluorescence-emitting compound that binds to the steroid hormone receptors in a solution containing the steroid hormone receptors. Then, measuring the fluorescence polarization of the solution. Subsequently, incubating the solution with at least one molecule that may compete with the compound for interaction with the steroid hormone receptors. Measuring the fluorescence polarization of the solution again. Finally, comparing the fluorescence polarization measurements to quantify any competitive interaction.

Method for quantitating competitive binding of molecules to proteins utilizing fluorescence polarization

A storable biotransformation mixture that can oxidize a drug, comprising a cytochrome P450 enzyme, an electron donor; and, a buffer solution. In a preferred embodiment the mixture also contains liposomes, cytochrome b.sub.5 and NADPH-P450 reductase as the electron donor. The mixture is storable at a temperature less than 8.degree. C. to for at least two weeks and is able to oxidize a drug after storage.

Growing community of inventors

Belleville, WI, United States of America

Robert G Lowery