Publication

Stepwise activation mechanism of the scramblase nhTMEM16 revealed by cryo-EM

Kalienkova, V., Mosina, V. C., Bryner, L., Oostergetel, G. T., Dutzler, R. & Paulino, C., 21-Feb-2019, In : eLife. 8, 27 p., 44364.

Research output: Contribution to journalArticleAcademicpeer-review

APA

Kalienkova, V., Mosina, V. C., Bryner, L., Oostergetel, G. T., Dutzler, R., & Paulino, C. (2019). Stepwise activation mechanism of the scramblase nhTMEM16 revealed by cryo-EM. eLife, 8, [44364]. https://doi.org/10.7554/eLife.44364

Author

Kalienkova, Valeria ; Mosina, Vanessa Clerico ; Bryner, Laura ; Oostergetel, Gert T. ; Dutzler, Raimund ; Paulino, Cristina. / Stepwise activation mechanism of the scramblase nhTMEM16 revealed by cryo-EM. In: eLife. 2019 ; Vol. 8.

Harvard

Kalienkova, V, Mosina, VC, Bryner, L, Oostergetel, GT, Dutzler, R & Paulino, C 2019, 'Stepwise activation mechanism of the scramblase nhTMEM16 revealed by cryo-EM', eLife, vol. 8, 44364. https://doi.org/10.7554/eLife.44364

Standard

Stepwise activation mechanism of the scramblase nhTMEM16 revealed by cryo-EM. / Kalienkova, Valeria; Mosina, Vanessa Clerico; Bryner, Laura; Oostergetel, Gert T.; Dutzler, Raimund; Paulino, Cristina.

In: eLife, Vol. 8, 44364, 21.02.2019.

Research output: Contribution to journalArticleAcademicpeer-review

Vancouver

Kalienkova V, Mosina VC, Bryner L, Oostergetel GT, Dutzler R, Paulino C. Stepwise activation mechanism of the scramblase nhTMEM16 revealed by cryo-EM. eLife. 2019 Feb 21;8. 44364. https://doi.org/10.7554/eLife.44364


BibTeX

@article{9e5c7f084f2140c9a553a32b8233cd21,
title = "Stepwise activation mechanism of the scramblase nhTMEM16 revealed by cryo-EM",
abstract = "Scramblases catalyze the movement of lipids between both leaflets of a bilayer. Whereas the X-ray structure of the protein nhTMEM16 has previously revealed the architecture of a Ca2+-dependent lipid scramblase, its regulation mechanism has remained elusive. Here, we have used cryo-electron microscopy and functional assays to address this question. Ca2+-bound and Ca2+-free conformations of nhTMEM16 in detergent and lipid nanodiscs illustrate the interactions with its environment and they reveal the conformational changes underlying its activation. In this process, Ca2+ binding induces a stepwise transition of the catalytic subunit cavity, converting a closed cavity that is shielded from the membrane in the absence of ligand, into a polar furrow that becomes accessible to lipid headgroups in the Ca2+-bound state. Additionally, our structures demonstrate how nhTMEM16 distorts the membrane at both entrances of the subunit cavity, thereby decreasing the energy barrier for lipid movement.",
keywords = "TMEM16A, PHOSPHATIDYLSERINE, VISUALIZATION, ORIENTATION, PROTEINS, EXPOSURE, CLONING, SYSTEM",
author = "Valeria Kalienkova and Mosina, {Vanessa Clerico} and Laura Bryner and Oostergetel, {Gert T.} and Raimund Dutzler and Cristina Paulino",
year = "2019",
month = "2",
day = "21",
doi = "10.7554/eLife.44364",
language = "English",
volume = "8",
journal = "eLife",
issn = "2050-084X",
publisher = "ELIFE SCIENCES PUBLICATIONS LTD",

}

RIS

TY - JOUR

T1 - Stepwise activation mechanism of the scramblase nhTMEM16 revealed by cryo-EM

AU - Kalienkova, Valeria

AU - Mosina, Vanessa Clerico

AU - Bryner, Laura

AU - Oostergetel, Gert T.

AU - Dutzler, Raimund

AU - Paulino, Cristina

PY - 2019/2/21

Y1 - 2019/2/21

N2 - Scramblases catalyze the movement of lipids between both leaflets of a bilayer. Whereas the X-ray structure of the protein nhTMEM16 has previously revealed the architecture of a Ca2+-dependent lipid scramblase, its regulation mechanism has remained elusive. Here, we have used cryo-electron microscopy and functional assays to address this question. Ca2+-bound and Ca2+-free conformations of nhTMEM16 in detergent and lipid nanodiscs illustrate the interactions with its environment and they reveal the conformational changes underlying its activation. In this process, Ca2+ binding induces a stepwise transition of the catalytic subunit cavity, converting a closed cavity that is shielded from the membrane in the absence of ligand, into a polar furrow that becomes accessible to lipid headgroups in the Ca2+-bound state. Additionally, our structures demonstrate how nhTMEM16 distorts the membrane at both entrances of the subunit cavity, thereby decreasing the energy barrier for lipid movement.

AB - Scramblases catalyze the movement of lipids between both leaflets of a bilayer. Whereas the X-ray structure of the protein nhTMEM16 has previously revealed the architecture of a Ca2+-dependent lipid scramblase, its regulation mechanism has remained elusive. Here, we have used cryo-electron microscopy and functional assays to address this question. Ca2+-bound and Ca2+-free conformations of nhTMEM16 in detergent and lipid nanodiscs illustrate the interactions with its environment and they reveal the conformational changes underlying its activation. In this process, Ca2+ binding induces a stepwise transition of the catalytic subunit cavity, converting a closed cavity that is shielded from the membrane in the absence of ligand, into a polar furrow that becomes accessible to lipid headgroups in the Ca2+-bound state. Additionally, our structures demonstrate how nhTMEM16 distorts the membrane at both entrances of the subunit cavity, thereby decreasing the energy barrier for lipid movement.

KW - TMEM16A

KW - PHOSPHATIDYLSERINE

KW - VISUALIZATION

KW - ORIENTATION

KW - PROTEINS

KW - EXPOSURE

KW - CLONING

KW - SYSTEM

U2 - 10.7554/eLife.44364

DO - 10.7554/eLife.44364

M3 - Article

VL - 8

JO - eLife

JF - eLife

SN - 2050-084X

M1 - 44364

ER -

ID: 95844308