Heterologously-expressed and Liposome-reconstituted Human Transient Receptor Potential Melastatin 4 Channel (TRPM4) is a Functional Tetramer

Scientific Reports
Maryrose ConstantineBoris Martinac

Abstract

Mutation, irregular expression and sustained activation of the Transient Receptor Potential Channel, type Melastatin 4 (TRPM4), have been linked to various cardiovascular diseases. However, much remains unknown about the structure of this important ion channel. Here, we have purified a heterologously expressed TRPM4-eGFP fusion protein and investigated the oligomeric state of TRPM4-eGFP in detergent micelles using crosslinking, native gel electrophoresis, multi-angle laser light scattering and electron microscopy. Our data indicate that TRPM4 is tetrameric, like other TRP channels studied to date. Furthermore, the functionality of liposome reconstituted TRPM4-eGFP was examined using electrophysiology. Single-channel recordings from TRPM4-eGFP proteoliposomes showed inhibition of the channel using Flufenamic acid, a well-established inhibitor of TRPM4, suggesting that the channels are functional upon reconstitution. Our characterisation of the oligomeric structure of TRPM4 and the ability to reconstitute functional channels in liposomes should facilitate future studies into the structure, function and pharmacology of this therapeutically relevant channel.

References

Mar 16, 2000·Trends in Neurosciences·C HarteneckG Schultz
Mar 21, 2002·Proceedings of the National Academy of Sciences of the United States of America·Boris Martinac, Owen P Hamill
Dec 5, 2003·Nature·David E Clapham
Aug 28, 2004·The Journal of Physiology·Bernd NiliusGuy Droogmans
Nov 20, 2004·Science·Pierre LaunayJean-Pierre Kinet
Apr 23, 2005·The Journal of Biological Chemistry·Bernd NiliusThomas Voets
May 10, 2005·The Journal of Physiology·Bernd NiliusThomas Voets
Jun 21, 2005·Biochemical and Biophysical Research Communications·Kazuhiro MioChikara Sato
Sep 28, 2005·The Journal of Biological Chemistry·Zheng ZhangEmily R Liman
May 9, 2006·Pflügers Archiv : European journal of physiology·Bernd Nilius, Rudi Vennekens
Feb 6, 2007·American Journal of Physiology. Heart and Circulatory Physiology·Nico H L KuijpersPeter A J Hilbers
Apr 11, 2007·Journal of Pharmacological Sciences·Hiromitsu MoritaJoseph E Brayden
Nov 6, 2007·Biophysical Journal·Michelle A DigmanEnrico Gratton
Mar 18, 2008·Progress in Biophysics and Molecular Biology·Joost H A FolgeringEric Honoré
Jul 16, 2008·Methods : a Companion to Methods in Enzymology·Dirk Jan SlotboomGuus B Erkens
Jun 9, 2009·Pharmacology & Therapeutics·Ryuji InoueYasuhiro Kawarabayashi
Jul 15, 2009·Journal of Cardiovascular Pharmacology·Jonathan A StiberPaul B Rosenberg
Sep 4, 2009·The Journal of Clinical Investigation·Martin KruseOlaf Pongs
Dec 23, 2009·Quarterly Reviews of Biophysics·Ramon LatorreSebastian Brauchi
Jan 2, 2010·The Journal of Biological Chemistry·Hideki ShigematsuKuniaki Nagayama
Jun 22, 2010·Circulation. Cardiovascular Genetics·Hui LiuPatrice Bouvagnet
Dec 22, 2010·The Journal of Physiology·Rudi Vennekens
Feb 4, 2011·Advances in Experimental Medicine and Biology·Minghui LiJian Yang
Feb 22, 2012·Nature·Bertrand CosteArdem Patapoutian
Apr 17, 2012·Circulation Journal : Official Journal of the Japanese Circulation Society·Tetsuo Minamino
May 16, 2012·Proceedings of the National Academy of Sciences of the United States of America·Takeshi NomuraBoris Martinac
Jul 4, 2012·Biochemical Pharmacology·Hugues AbrielJean-Sébastien Rougier
Feb 19, 2013·Journal of Molecular and Cellular Cardiology·Christophe SimardRomain Guinamard
Feb 28, 2013·Journal of Cellular and Molecular Medicine·Ken TakahashiKeiji Naruse
Feb 28, 2013·Pflügers Archiv : European journal of physiology·Scott Earley
Jul 28, 2013·Biochimica Et Biophysica Acta·Boris Martinac
Jan 18, 2014·British Journal of Pharmacology·R GuinamardC A Del Negro
Apr 12, 2014·Current Opinion in Pharmacology·Martin Kruse, Olaf Pongs
Aug 8, 2014·Journal of Cerebral Blood Flow and Metabolism : Official Journal of the International Society of Cerebral Blood Flow and Metabolism·Yao LiJoseph E Brayden
Dec 3, 2014·BMB Reports·Chang-Hoon ChoJae-Yong Park
Dec 3, 2014·Journal of Molecular Biology·Jennifer A Thomas, Christopher G Tate
Dec 17, 2014·Proceedings of the National Academy of Sciences of the United States of America·Jonas M DörrJ Antoinette Killian
Mar 5, 2015·Proceedings of the National Academy of Sciences of the United States of America·Jinfeng TengChing Kung

❮ Previous
Next ❯

Citations

Dec 7, 2017·Nature·Paige A WinklerWei Lü
Nov 15, 2019·Journal of Cell Science·Yury A NikolaevBoris Martinac
May 27, 2017·The Biochemical Journal·Ioli MitsouJohn R Couchman
Aug 9, 2020·Progress in Biophysics and Molecular Biology·Yang GuoOliver Friedrich

❮ Previous
Next ❯

Methods Mentioned

BETA
electron microscopy
confocal microscopy
size-exclusion chromatography
size-exclusion purification
Electrophoresis
size
static light scattering
electron
Fluorescence lifetime
PCR

Software Mentioned

Multi Gauge
SimFCS
Clampfit
QuB
GraphPad Prism
pClamp
ASTRA

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.