Understanding the molecular mechanism of TRP channel activation/inhibition by structural analysis
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Case Western Reserve University School of Graduate Studies / OhioLINK
2018
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| Online Access: | http://rave.ohiolink.edu/etdc/view?acc_num=case1533499522272657 |
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ndltd-OhioLink-oai-etd.ohiolink.edu-case15334995222726572021-08-03T07:08:04Z Understanding the molecular mechanism of TRP channel activation/inhibition by structural analysis Samanta, Amrita Biophysics Transient Receptor Potential (TRP) superfamily of ion channels are evolutionarily conserved integral membrane proteins. The 28 members of the mammalian TRP superfamily are cation permeable channels which are grouped into six subfamilies based on their sequence homology. This superfamily of ion channels is involved in a vast array of physiological and pathophysiological processes including cancer, neuropathic pain, cardiovascular disease, inflammation, diabetes and polycystic kidney disease. Therefore, the study of these channels is imperative to our better understanding of subcellular biochemistry. The latter half of the 20th century marked the discovery of TRP channels and a lot is known about their physiological characteristics and functions. But until the last couple of years, not much was known about their structure and molecular mechanisms underlying their biophysical and physiological properties. Here I utilized our ability to heterologously express and purify full length, stable and functional mammalian TRP channels to gain insight into the molecular mechanism of gating of two of the TRP channels: TRPV2 and TRPA1. Using cannabidiol (CBD), an activator of TRPV2 channel, we tested the hypothesis that the pore turret region of the channel is important for opening of its ion conduction pore and could also capture the open state structure of TRPV2 using cryo electron microscopy (cryo-EM) revealing the gating mechanism of this channel during activation. In case of TRPA1, it was known that certain cysteine residues are critical for channel activation with electrophilic ligands, however the mechanism of channel gating was unknown. Moreover, not much was known about effect of non-electrophilic modulators on this channel. In second part of this study I illustrated the regions of TRPA1 ion channel that undergoes ligand induced conformational changes using limited proteolysis coupled with mass spectrometry. The results showed that in case of TRPA1, irrespective of the ligand, the linker region and the N-terminal ankyrin repeats experience conformational changes leading to activation or inhibition of the channel. 2018-08-31 English text Case Western Reserve University School of Graduate Studies / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=case1533499522272657 http://rave.ohiolink.edu/etdc/view?acc_num=case1533499522272657 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws. |
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NDLTD |
| language |
English |
| sources |
NDLTD |
| topic |
Biophysics |
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Biophysics Samanta, Amrita Understanding the molecular mechanism of TRP channel activation/inhibition by structural analysis |
| author |
Samanta, Amrita |
| author_facet |
Samanta, Amrita |
| author_sort |
Samanta, Amrita |
| title |
Understanding the molecular mechanism of TRP channel activation/inhibition by structural analysis |
| title_short |
Understanding the molecular mechanism of TRP channel activation/inhibition by structural analysis |
| title_full |
Understanding the molecular mechanism of TRP channel activation/inhibition by structural analysis |
| title_fullStr |
Understanding the molecular mechanism of TRP channel activation/inhibition by structural analysis |
| title_full_unstemmed |
Understanding the molecular mechanism of TRP channel activation/inhibition by structural analysis |
| title_sort |
understanding the molecular mechanism of trp channel activation/inhibition by structural analysis |
| publisher |
Case Western Reserve University School of Graduate Studies / OhioLINK |
| publishDate |
2018 |
| url |
http://rave.ohiolink.edu/etdc/view?acc_num=case1533499522272657 |
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AT samantaamrita understandingthemolecularmechanismoftrpchannelactivationinhibitionbystructuralanalysis |
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