video: Using cryo-EM, scientists at Columbia University have revealed structural changes in the temperature-sensing molecule TRPV3 when activated by heat. view more
Credit: Sobolevsky Lab, Columbia University Vagelos College of Physicians and Surgeons
NEW YORK, NY (October 21, 2019)--Columbia University researchers have captured new detailed images of a temperature-sensing molecule in its open, intermediate, and closed states. The findings will help us understand the mechanics of hot, warm, cool, and cold sensation and accelerate the development of drugs for variety of conditions, including inflammatory skin disease, itch, and pain.
The findings were published online in Nature Structural and Molecular Biology.
Researchers have known for more than a century that temperature and pain perception are controlled by specialized sensory neurons. In the late 1990s, scientists discovered a superfamily of molecules, called transient receptor potential (TRP) channels, inside these neurons. Of the 28 known TRP channels, 11 are highly sensitive to changes in temperature. These "thermoTRPs" open and close in response to temperature fluctuations, allowing ions to pass through and transmit signals to the central nervous system.
"ThermoTRPs act as biological thermometers, allowing organisms to sense temperatures in the entire physiological range, from noxious cold to noxious heat," says study leader Alexander I. Sobolevsky, PhD, associate professor of biochemistry and molecular biophysics at Columbia University Vagelos College of Physicians and Surgeons. "How these channels sense temperature and then subsequently undergo changes at the molecular level has remained a puzzle."
An important breakthrough in the field came in 2013 when a technique called cryo-electron microscopy (cryo-EM) was first used to image representatives of the TRP channel family. Since then, almost every type of TRP channel has been imaged with this technique. But scientists were unable to capture cryo-EM images of thermoTRP channels at different temperatures--particularly high temperatures, where heat-sensitive channels are activated.
In the new study, Sobolevsky's lab identified a mutation in one of these channels, TRPV3, that increases its sensitivity to temperature. By exploiting this mutation, his team was able to fix the channel in open, sensitized, and closed positions--an essential step in understanding a protein's structure using cryo-EM imaging. The resultant images revealed the structural changes that happen in TRPV3 once it is activated by heat.
"These structural changes seem to originate from the ion-conducting membrane portion of TRPV3 that senses temperature through its interaction with the surrounding membrane lipids," says Appu K. Singh, PhD, an associate research scientist in biochemistry and molecular biophysics at Columbia University and first author of the paper. "Further studies are needed to identify the temperature sensor of these channels more precisely."
TRPV3 is mainly expressed in skin cells. The channel plays a role in a variety of physiological functions, including temperature sensation, nociception (the nervous system's response to certain harmful or potentially harmful stimuli), itch, maintenance of the skin barrier, wound healing, and hair growth. TRPV3 is also associated with numerous human skin diseases, such as atopic dermatitis, psoriasis, and rosacea. Studies show that mice lacking TRPV3 are unable to sense warmer temperatures.
"Our structures not only can serve as a springboard for studies of the biophysical principles of ion channel temperature activation, but also as templates for the design of drugs for a variety of conditions that affect the skin," says Sobolevsky.
In a separate study published in Nature Communications, Sobolevsky's team used cryo-EM to determine the structure of crTRP1, a thermoTRP found in Chlamydomonas reinhardtii, a type of algae. This is the first time the structure of a thermoTRP has been described in a micro-organism.
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Video: https://youtu.be/Qp_CjeUAytA
The paper is titled, "Structural basis of temperature sensation by the TRP channel TRPV3." The other contributors are: Luke L. McGoldrick (Columbia University Irving Medical Center), Lusine Demirkhanyan (University of Illinois College of Medicine), Merfilius Leslie (University of Illinois), and Eleonora Zakharian (University of Illinois).
The study was supported by grants from the National Institutes of Health (F31 CA232391-01, R01 CA206573, R01 5 NS083660, R01 NS107253, and GM103310), National Science Foundation, Irma T. Hirschl Career Scientist Award. Simons Foundation, and NYSTAR (the Empire State Development's Division of Science, Technology and Innovation).
The authors declare no financial or other conflicts of interest.
Columbia University Irving Medical Center provides international leadership in basic, preclinical, and clinical research; medical and health sciences education; and patient care. The medical center trains future leaders and includes the dedicated work of many physicians, scientists, public health professionals, dentists, and nurses at the Vagelos College of Physicians and Surgeons, the Mailman School of Public Health, the College of Dental Medicine, the School of Nursing, the biomedical departments of the Graduate School of Arts and Sciences, and allied research centers and institutions. Columbia University Irving Medical Center is home to the largest medical research enterprise in New York City and State and one of the largest faculty medical practices in the Northeast. For more information, visit cuimc.columbia.edu or columbiadoctors.org.
Journal
Nature Structural & Molecular Biology