Drug Activity Observed in Solitary Molecule While in Motion


NEW YORK — Weill Cornell Medical College researchers have developed a new technology that allows them to directly visualize functional motions of individual enzymes in nearly real time. The new development demonstrates for the first time how molecular movements are affected by antibiotic binding. The findings provide an important new perspective into biochemical processes that may lead to the development of new drug therapies.

The study is supported by funds from the US National Institute of General Medical Sciences and published in a recent issue of Nature Chemical Biology.

“Understanding molecular movements is important because enzyme function hinges on motion,” explains Dr. Scott Blanchard, senior author and associate professor of physiology and biophysics at Weill Cornell Medical College in New York City. “To observe the molecule, we are decorating it with fluorescent markers, called fluorophores, that make it glow.”

The fluorophores are attached to the biomolecule and are designed to exchange energy with each other in a way that accurately reports on the distance between them, like a molecular global positioning system. This process is called fluorescence resonance energy transfer (FRET). When applied to the study of single-molecules (smFRET), one can actually use this technique to monitor changes in the structure of individual enzymes as they function.

Traditionally, in order to understand how drugs affect enzymes, researchers have measured changes in the rate at which an enzyme generates product, which often requires a great deal of starting material. The new single-molecule approach provides the ability to observe enzyme function from the perspective of motion, and how such motions are influenced by the presence of substrates or drug compounds.

In the current study, Dr. Blanchard and his team investigated whether the binding of aminoglycoside-class antibiotics — an important family of clinically useful small-molecule compounds — affects how the ribosome moves. The ribosome, one of the largest and most essential molecular machines in the cell, is the target of almost half of all known antibiotics currently in use.

The aminoglycosides, while highly effective, tend to be toxic. Dr. Blanchard’s goal was to explore the relationship between aminoglycoside activities and ribosome movements and to search for compounds with more potent activities but that have fewer side effects.

While the approach has many advantages, one of the most valuable is that it is “green,” explains Dr. Blanchard. As implied by its name, single-molecule methods are characterized by a greatly reduced demand for biological material. Consequently, less human and capital costs go into large-scale sample preparations. In principle, the researchers believe the single-molecule technique may one day be engineered to require a million times less starting material than is required by traditional drug screening methods.

“In addition to this material advantage, the information content of the single-molecule approach is greater, increasing the cost effectiveness of each experiment,” explains Dr. Blanchard. “Our challenge now is to understand whether the approach is generalizable to other enzyme systems where an understanding of its regulation and the mechanism of drug action are lacking.”

Co-authors of the study include Michael Feldman, Daniel Terry and Roger Altman, all from Weill Cornell.

Weill Cornell Medical College

Weill Cornell Medical College, Cornell University’s medical school located in New York City, is committed to excellence in research, teaching, patient care and the advancement of the art and science of medicine, locally, nationally and globally. Physicians and scientists of Weill Cornell Medical College are engaged in cutting-edge research from bench to bedside, aimed at unlocking mysteries of the human body in health and sickness and toward developing new treatments and prevention strategies. In its commitment to global health and education, Weill Cornell has a strong presence in places such as Qatar, Tanzania, Haiti, Brazil, Austria and Turkey. Through the historic Weill Cornell Medical College in Qatar, the Medical College is the first in the U.S. to offer its M.D. degree overseas. Weill Cornell is the birthplace of many medical advances — including the development of the Pap test for cervical cancer, the synthesis of penicillin, the first successful embryo-biopsy pregnancy and birth in the U.S., the first clinical trial of gene therapy for Parkinson’s disease, and most recently, the world’s first successful use of deep brain stimulation to treat a minimally conscious brain-injured patient. Weill Cornell Medical College is affiliated with NewYork-Presbyterian Hospital, where its faculty provides comprehensive patient care at NewYork-Presbyterian Hospital/Weill Cornell Medical Center. The Medical College is also affiliated with the Methodist Hospital in Houston, making Weill Cornell one of only two medical colleges in the country affiliated with two U.S.News & World Report Honor Roll hospitals. For more information, visit www.med.cornell.edu.

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Andrew Klein

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