Heterologous expression and purification of an active human TRPV3 ion channel

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Heterologous expression and purification of an active human TRPV3 ion channel. / Kol, S.; Braun, Christian Seeberg; Thiel, G.; Doyle, D.A.; Sundström, M.; Gourdon, Pontus Emanuel; Nissen, Poul.

In: The F E B S Journal (Online), Vol. 280, No. 23, 01.12.2013, p. 6010-6021.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Kol, S, Braun, CS, Thiel, G, Doyle, DA, Sundström, M, Gourdon, PE & Nissen, P 2013, 'Heterologous expression and purification of an active human TRPV3 ion channel', The F E B S Journal (Online), vol. 280, no. 23, pp. 6010-6021. https://doi.org/10.1111/febs.12520

APA

Kol, S., Braun, C. S., Thiel, G., Doyle, D. A., Sundström, M., Gourdon, P. E., & Nissen, P. (2013). Heterologous expression and purification of an active human TRPV3 ion channel. The F E B S Journal (Online), 280(23), 6010-6021. https://doi.org/10.1111/febs.12520

Vancouver

Kol S, Braun CS, Thiel G, Doyle DA, Sundström M, Gourdon PE et al. Heterologous expression and purification of an active human TRPV3 ion channel. The F E B S Journal (Online). 2013 Dec 1;280(23):6010-6021. https://doi.org/10.1111/febs.12520

Author

Kol, S. ; Braun, Christian Seeberg ; Thiel, G. ; Doyle, D.A. ; Sundström, M. ; Gourdon, Pontus Emanuel ; Nissen, Poul. / Heterologous expression and purification of an active human TRPV3 ion channel. In: The F E B S Journal (Online). 2013 ; Vol. 280, No. 23. pp. 6010-6021.

Bibtex

@article{3ed31e0827814d33844c3cd74a270b52,

title = "Heterologous expression and purification of an active human TRPV3 ion channel",

abstract = "The transient receptor potential vanilloid 3 (TRPV3) cation channel is widely expressed in human tissues and has been shown to be activated by mild temperatures or chemical ligands. In spite of great progress in the TRP-channel characterization, very little is known about their structure and interactions with other proteins at the atomic level. This is mainly caused by difficulties in obtaining functionally active samples of high homogeneity. Here, we report on the high-level Escherichia coli expression of the human TRPV3 channel, for which no structural information has been reported to date. We selected a suitable detergent and buffer system using analytical size-exclusion chromatography and a thermal stability assay. We demonstrate that the recombinant purified protein contains high α-helical content and migrates as dimers and tetramers on native PAGE. Furthermore, the purified channel also retains its current inducing activity, as shown by electrophysiology experiments. The ability to produce the TRPV3 channel heterologously will aid future functional and structural studies.",

author = "S. Kol and Braun, {Christian Seeberg} and G. Thiel and D.A. Doyle and M. Sundstr{\"o}m and Gourdon, {Pontus Emanuel} and Poul Nissen",

year = "2013",

month = dec,

day = "1",

doi = "10.1111/febs.12520",

language = "English",

volume = "280",

pages = "6010--6021",

journal = "The F E B S Journal (Online)",

issn = "1742-4658",

publisher = "Wiley-Blackwell",

number = "23",

}

RIS

TY - JOUR

T1 - Heterologous expression and purification of an active human TRPV3 ion channel

AU - Kol, S.

AU - Braun, Christian Seeberg

AU - Thiel, G.

AU - Doyle, D.A.

AU - Sundström, M.

AU - Gourdon, Pontus Emanuel

AU - Nissen, Poul

PY - 2013/12/1

Y1 - 2013/12/1

N2 - The transient receptor potential vanilloid 3 (TRPV3) cation channel is widely expressed in human tissues and has been shown to be activated by mild temperatures or chemical ligands. In spite of great progress in the TRP-channel characterization, very little is known about their structure and interactions with other proteins at the atomic level. This is mainly caused by difficulties in obtaining functionally active samples of high homogeneity. Here, we report on the high-level Escherichia coli expression of the human TRPV3 channel, for which no structural information has been reported to date. We selected a suitable detergent and buffer system using analytical size-exclusion chromatography and a thermal stability assay. We demonstrate that the recombinant purified protein contains high α-helical content and migrates as dimers and tetramers on native PAGE. Furthermore, the purified channel also retains its current inducing activity, as shown by electrophysiology experiments. The ability to produce the TRPV3 channel heterologously will aid future functional and structural studies.

AB - The transient receptor potential vanilloid 3 (TRPV3) cation channel is widely expressed in human tissues and has been shown to be activated by mild temperatures or chemical ligands. In spite of great progress in the TRP-channel characterization, very little is known about their structure and interactions with other proteins at the atomic level. This is mainly caused by difficulties in obtaining functionally active samples of high homogeneity. Here, we report on the high-level Escherichia coli expression of the human TRPV3 channel, for which no structural information has been reported to date. We selected a suitable detergent and buffer system using analytical size-exclusion chromatography and a thermal stability assay. We demonstrate that the recombinant purified protein contains high α-helical content and migrates as dimers and tetramers on native PAGE. Furthermore, the purified channel also retains its current inducing activity, as shown by electrophysiology experiments. The ability to produce the TRPV3 channel heterologously will aid future functional and structural studies.

UR - http://www.scopus.com/inward/record.url?scp=84888305651&partnerID=8YFLogxK

U2 - 10.1111/febs.12520

DO - 10.1111/febs.12520

M3 - Journal article

C2 - 24028292

AN - SCOPUS:84888305651

VL - 280

SP - 6010

EP - 6021

JO - The F E B S Journal (Online)

JF - The F E B S Journal (Online)

SN - 1742-4658

IS - 23

ER -

ID: 88649471