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Quick, easy, color-coded technique will speed identification of drugs for diseases

John Hopkins researchers have developed the first color-coded tracking system to see how receptors on the surface of a living cell transmit signals to the cell's interior and regulate a wide range of biological processes. The technology, described in the March 23 issue of Science, should significantly speed up the search for drugs needed to treat heart disease, cancer, asthma and other ailments.

"For the first time, we can actually see the activation of GPCRs, receptors that set off many cascades in a cell," says Peter Devreotes, Ph.D., director of the Department of Cell Biology and Anatomy at Hopkins and principal investigator of the study. "Instead of having to screen for effective drugs by estimating their secondary chemical effects, we can see the action directly on the receptor."

A vast family of GPCRs, or G-protein coupled receptors, trigger changes within cells when set off by hormones, neurotransmitters, chemoattractants and other proteins. Researchers have known that an inactivated GPCR holds onto three G-protein pieces (alpha, beta and gamma) that dangle into the interior of the cell. When a molecule activates and binds to a receptor's "docking bay" on the outside of the cell, the receptor causes the three pieces to drift apart.

Devreotes and his colleagues created analog proteins that mimic the alpha and beta forms and tagged them with fluorescent proteins. The researchers hoped these markers would be so close together that a process known as FRET (fluorescence resonance energy transfer) would occur and they would fluoresce yellow. If the receptors were activated, these tags would drift apart and they would show up as cyan under the microscope.

Using a one-celled amoeba called Dictyostelium, whose GPCRs serve as docking bays for a chemical known as cyclic AMP (cAMP), the researchers added alpha and beta analogs to the genome of the amoeba and then exposed the cells to cAMP. When the GPCR was inactive, it emitted yellow
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Contact: Kate O'Rourke
korourke@jhmi.edu
410-955-8665
Johns Hopkins Medical Institutions
22-Mar-2001


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