scholarly journals Genetic variants affecting human TRPA1 or TRPM8 structure can be classified in vitro as ‘well expressed’, ‘poorly expressed’ or ‘salvageable’

2015 ◽  
Vol 35 (5) ◽  
Author(s):  
Kevin Morgan ◽  
Laura Rachel Sadofsky ◽  
Alyn Hugh Morice
Keyword(s):  
Genetic Variants ◽  
Transient Receptor Potential ◽  
Kinase Inhibitor ◽  
Receptor Potential ◽  
Transient Receptor Potential Channels ◽  
Human Embryonic Kidney ◽  
Potential Channels ◽  
Pre Treatment ◽  
Transient Receptor

Genetic variants of human transient receptor potential channels A1 and M8 expressed in human embryonic kidney HEK293 and SH-SY5Y cells were assayed using Ca2+ signalling. TRPA1 Y69C responded well. Poorly expressed variant signalling was enhanced by pre-treatment with tyrosine kinase inhibitor PP2 or Zn2+.

scholarly journals The Membrane Proximal Domain of TRPV1 and TRPV2 Channels Mediates Protein–Protein Interactions and Lipid Binding In Vitro

2019 ◽  
Vol 20 (3) ◽  
pp. 682 ◽  
Author(s):  
Pau Doñate-Macián ◽  
Elena Álvarez-Marimon ◽  
Francesc Sepulcre ◽  
José Vázquez-Ibar ◽  
Alex Perálvarez-Marín
Keyword(s):  
Protein Interactions ◽  
Transient Receptor Potential ◽  
Receptor Potential ◽  
Lipid Binding ◽  
Transient Receptor Potential Channels ◽  
Trpv Channels ◽  
Potential Channels ◽  
Transient Receptor ◽  
Lipid Protein Interactions

Constitutive or regulated membrane protein trafficking is a key cell biology process. Transient receptor potential channels are somatosensory proteins in charge of detecting several physical and chemical stimuli, thus requiring fine vesicular trafficking. The membrane proximal or pre-S1 domain (MPD) is a highly conserved domain in transient receptor potential channels from the vanilloid (TRPV) subfamily. MPD shows traits corresponding to protein-protein and lipid-protein interactions, and protein regulatory regions. We have expressed MPD of TRPV1 and TRPV2 as green fluorescente protein (GFP)-fusion proteins to perform an in vitro biochemical and biophysical characterization. Pull-down experiments indicate that MPD recognizes and binds Soluble N-ethylmaleimide-sensitive factor Attachment Protein Receptors (SNARE). Synchrotron radiation scattering experiments show that this domain does not self-oligomerize. MPD interacts with phosphatidic acid (PA), a metabolite of the phospholipase D (PLD) pathway, in a specific manner as shown by lipid strips and Trp fluorescence quenching experiments. We show for the first time, to the best of our knowledge, the binding to PA of an N-terminus domain in TRPV channels. The presence of a PA binding domain in TRPV channels argues for putative PLD regulation. Findings in this study open new perspectives to understand the regulated and constitutive trafficking of TRPV channels exerted by protein-protein and lipid-protein interactions.

scholarly journals FeRIC-based magnetogenetics: evaluation of methods and protocols in in vitro models

2020 ◽  
Author(s):  
Miriam Hernández-Morales ◽  
Victor Han ◽  
Richard H Kramer ◽  
Chunlei Liu
Keyword(s):  
Transient Receptor Potential ◽  
Receptor Potential ◽  
Functional Expression ◽  
In Vitro Models ◽  
Transient Receptor Potential Channels ◽  
Iron Level ◽  
Ferritin Iron ◽  
Potential Channels ◽  
Transient Receptor

AbstractFeRIC (Ferritin iron Redistribution to Ion Channels) is a magnetogenetic technique that uses radiofrequency (RF) waves to activate the transient receptor potential channels, such as TRPV1 and TRPV4, coupled to cellular ferritins. In cells expressing ferritin-tagged TRPV, RF stimulation increases the cytosolic Ca2+ levels via a biochemical pathway. The interaction between RF and ferritin increases the free cytosolic iron level that in turn, triggers chemical reactions producing reactive oxygen species and oxidized lipids that activate the ferritin-tagged TRPV. In this pathway, it is expected that experimental factors that disturb the ferritin expression, the ferritin iron load, the TRPV functional expression, or the cellular redox state will impact the RF efficacy to activate ferritin-tagged TRPV. Here, three in vitro protocols were compared for using FeRIC to remotely activate ferritin-tagged TRPV. Further, several experimental factors were examined that either enhance or abolish the RF control of ferritin-tagged TRPV. The findings may help establish reproducible magnetogenetic experimental protocols.

scholarly journals Effects of Ginger and its Pungent Constituents on Transient Receptor Potential Channels

2017 ◽  
Vol 112 (3) ◽  
pp. 250a
Author(s):  
Young-Soo Kim ◽  
Chan Sik Hong ◽  
Sang Weon Lee ◽  
Joo Hyun Nam ◽  
Byung Joo Kim
Keyword(s):  
Transient Receptor Potential ◽  
Receptor Potential ◽  
Transient Receptor Potential Channels ◽  
Potential Channels ◽  
Transient Receptor

What Evolutionary Evidence Implies About the Identity of the Mechanoelectrical Couplers in Vascular Smooth Muscle Cells

Physiology ◽  
2021 ◽  
Vol 36 (5) ◽  
pp. 292-306
Author(s):  
Heather A. Drummond
Keyword(s):  
Transient Receptor Potential ◽  
Molecular Model ◽  
Receptor Potential ◽  
Transient Receptor Potential Channels ◽  
Potential Channels ◽  
Transient Receptor ◽  
Paradigm Shifting ◽  
G Protein Coupled ◽  
Evolutionary Role

Loss of pressure-induced vasoconstriction increases susceptibility to renal and cerebral vascular injury. Favored paradigms underlying initiation of the response include transient receptor potential channels coupled to G protein-coupled receptors or integrins as transducers. Degenerin channels may also mediate the response. This review addresses the 1) evolutionary role of these molecules in mechanosensing, 2) limitations to identifying mechanosensitive molecules, and 3) paradigm shifting molecular model for a VSMC mechanosensor.

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