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Development of a High-Throughput Assay to Measure Histidine Decarboxylase Activity

Department of Medicinal Chemistry, Biologics and Biomolecular Sciences,Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT

Lori Patnaude

Department of Medicinal Chemistry, Biologics and Biomolecular Sciences,Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT

Jerry Hopkins

Department of Medicinal Chemistry and Cardiovascular Disease, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT

Joey Studts

Department of Medicinal Chemistry and Cardiovascular Disease, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT

Elda Gautschi

Department of Medicinal Chemistry and Cardiovascular Disease, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT

Anthony Shrutkowski

Department of Medicinal Chemistry and Cardiovascular Disease, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT

Anthony Kronkaitis

Department of Medicinal Chemistry and Cardiovascular Disease, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT

Martha Brown

Department of Medicinal Chemistry and Cardiovascular Disease, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT

Alisa Kabcenell

Department of Medicinal Chemistry, Research and Development, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT

Daniel Rajotte

Department of Medicinal Chemistry, Research and Development, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT

Histamine is a well-known mediator of allergic, inflammatory, and neurological responses. More recent studies suggest a role for histamine and its receptors in a wide range of biological processes, including T-cell maturation and bone remodeling. Histamine serum levels are regulated mainly by the activity of the histamine-synthesizing enzyme histidine decarboxylase (HDC). Despite the importance of this enzyme in many physiological processes, very few potent HDC inhibitors have been identified. HDC assays suitable for high-throughput screening have not been reported. The authors describe the development of a fluorescence polarization assay to measure HDC enzymatic activity. They used a fluorescein-histamine probe that binds with high affinity to an antihistamine antibody for detection. Importantly, they show that probe binding is fully competed by histamine, but no competition by the HDC substrate histidine was observed. The automated assay was performed in a total volume of 60 µL, had an assay window of 80 to 100 mP, and had a Z' factor of 0.6 to 0.7. This assay provides new tools to study HDC activity and pharmacological modulation of histamine levels.

Key Words: histidine decarboxylase • histamine • screening • fluorescence polarization

This version was published on October 1, 2006

Journal of Biomolecular Screening, Vol. 11, No. 7, 816-821 (2006)
DOI: 10.1177/1087057106290803


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