Imaging redox metabolism in living cells

MEDI 278

Dominic J. Yee1, Vojtech Balsanek1, David Bauman2, Trevor M Penning3, and Dalibor Sames, sames@chem.columbia.edu1. (1) Department of Chemistry, Columbia University, 3000 Broadway, New York, NY 10027, (2) Department of Biochemistry & Biophysics, University of Pennsylvania, School of Medicine, Philadelphia, PA 19104, (3) Center of Excellence in Environmental Toxicology, Department of Pharmacology, University of Pennsylvania School of Medicine, 135 John Morgan Building, 3620 Hamilton Walk, Philadelphia, PA
An assessment of enzymatic activity in living systems currently relies on discontinuous and destructive assays. Immunoblotting and real-time RT-PCR methods quantify protein and RNA levels by cellular RNA and protein extraction. Meanwhile, natural substrate metabolism is commonly determined by radiochemical chromatagraphy. We introduce new small molecule fluorogenic reporter substrates for determining hydroxysteroid dehydrogenase (HSD) and monoamine oxidase (MAO) activity in living cells. The metabolic indicators were discovered through two libraries designed based on a reversible and irreversible fluorogenic reporting motif. We have demonstrated the utility of the optical reporters for direct and continuous readout of expression, activity, and inhibition of these key physiological enzymes in living cells.

 

Medicinal Chemistry Award Symposium
9:00 AM-12:10 PM, Tuesday, 12 September 2006 Moscone Center -- Room 102, Oral

Division of Medicinal Chemistry

The 232nd ACS National Meeting, San Francisco, CA, September 10-14, 2006