Saposin-Lipoprotein Scaffolds for Structure Determination of Membrane Transporters

Publikation: Bidrag til bog/antologi/rapport/proceedingBidrag til bog/antologiForskningpeer review

Standard

Saposin-Lipoprotein Scaffolds for Structure Determination of Membrane Transporters. / Lyons, Joseph A.; Bøggild, Andreas; Nissen, Poul; Frauenfeld, Jens.

Methods in Enzymology: A Structure-Function Toolbox for Membrane Transporter and Channels. red. / Christine Ziegler. Bind 594 Academic Press, 2017. s. 85-99 (Methods in Enzymology, Bind 594).

Publikation: Bidrag til bog/antologi/rapport/proceedingBidrag til bog/antologiForskningpeer review

Harvard

Lyons, JA, Bøggild, A, Nissen, P & Frauenfeld, J 2017, Saposin-Lipoprotein Scaffolds for Structure Determination of Membrane Transporters. i C Ziegler (red.), Methods in Enzymology: A Structure-Function Toolbox for Membrane Transporter and Channels. bind 594, Academic Press, Methods in Enzymology, bind 594, s. 85-99. https://doi.org/10.1016/bs.mie.2017.06.035

APA

Lyons, J. A., Bøggild, A., Nissen, P., & Frauenfeld, J. (2017). Saposin-Lipoprotein Scaffolds for Structure Determination of Membrane Transporters. I C. Ziegler (red.), Methods in Enzymology: A Structure-Function Toolbox for Membrane Transporter and Channels (Bind 594, s. 85-99). Academic Press. Methods in Enzymology, Bind. 594 https://doi.org/10.1016/bs.mie.2017.06.035

CBE

Lyons JA, Bøggild A, Nissen P, Frauenfeld J. 2017. Saposin-Lipoprotein Scaffolds for Structure Determination of Membrane Transporters. Ziegler C, red. I Methods in Enzymology: A Structure-Function Toolbox for Membrane Transporter and Channels. Academic Press. s. 85-99. (Methods in Enzymology, Bind 594). https://doi.org/10.1016/bs.mie.2017.06.035

MLA

Lyons, Joseph A. o.a.. "Saposin-Lipoprotein Scaffolds for Structure Determination of Membrane Transporters". Ziegler, Christine (redaktører). Methods in Enzymology: A Structure-Function Toolbox for Membrane Transporter and Channels. Kapitel 3, Academic Press. (Methods in Enzymology, Bind 594). 2017, 85-99. https://doi.org/10.1016/bs.mie.2017.06.035

Vancouver

Lyons JA, Bøggild A, Nissen P, Frauenfeld J. Saposin-Lipoprotein Scaffolds for Structure Determination of Membrane Transporters. I Ziegler C, red., Methods in Enzymology: A Structure-Function Toolbox for Membrane Transporter and Channels. Bind 594. Academic Press. 2017. s. 85-99. (Methods in Enzymology, Bind 594). https://doi.org/10.1016/bs.mie.2017.06.035

Author

Lyons, Joseph A. ; Bøggild, Andreas ; Nissen, Poul ; Frauenfeld, Jens. / Saposin-Lipoprotein Scaffolds for Structure Determination of Membrane Transporters. Methods in Enzymology: A Structure-Function Toolbox for Membrane Transporter and Channels. red. / Christine Ziegler. Bind 594 Academic Press, 2017. s. 85-99 (Methods in Enzymology, Bind 594).

Bibtex

@inbook{198c168b5c8c4ce49a0f407924a6f39a,
title = "Saposin-Lipoprotein Scaffolds for Structure Determination of Membrane Transporters",
abstract = "Membrane proteins depend on their natural lipid environment for function, which makes them more difficult to study in isolation. A number of approaches that mimic the lipid bilayer of biological membranes have been described (nanodiscs, SMALPs), enabling novel ways to assay activity and elucidate structures of this important class of proteins. More recently, the use of saposin A, a protein that is involved in lipid transport, to form Salipro (saposin–lipid–protein) complexes was demonstrated for a range of membrane protein targets (Frauenfeld et al., 2016). The method is fast and requires few resources. The saposin–lipid–scaffold adapts to various sizes of transmembrane regions during self-assembly, forming a minimal lipid nanoparticle. This results in the formation of a well-defined membrane protein–lipid complex, which is desirable for structural characterization. Here, we describe a protocol to reconstitute the sarco-endoplasmic reticulum calcium ATPase (SERCA) into Salipro nanoparticles. The complex formation is analyzed using negative stain electron microscopy (EM), allowing to quickly determine an initial structure of the membrane protein and to evaluate sample conditions for structural studies using single-particle cryo-EM in a detergent-free environment.",
keywords = "Cryo-EM, Detergent, Detergent-free, Electron microscopy, Lipids, Membrane protein structure, P-type ATPases, Salipro, Saposin, SERCA",
author = "Lyons, {Joseph A.} and Andreas B{\o}ggild and Poul Nissen and Jens Frauenfeld",
year = "2017",
doi = "10.1016/bs.mie.2017.06.035",
language = "English",
isbn = "978-0-12-812353-9",
volume = "594",
series = "Methods in Enzymology",
publisher = "Academic Press",
pages = "85--99",
editor = "Ziegler, {Christine }",
booktitle = "Methods in Enzymology",

}

RIS

TY - CHAP

T1 - Saposin-Lipoprotein Scaffolds for Structure Determination of Membrane Transporters

AU - Lyons, Joseph A.

AU - Bøggild, Andreas

AU - Nissen, Poul

AU - Frauenfeld, Jens

PY - 2017

Y1 - 2017

N2 - Membrane proteins depend on their natural lipid environment for function, which makes them more difficult to study in isolation. A number of approaches that mimic the lipid bilayer of biological membranes have been described (nanodiscs, SMALPs), enabling novel ways to assay activity and elucidate structures of this important class of proteins. More recently, the use of saposin A, a protein that is involved in lipid transport, to form Salipro (saposin–lipid–protein) complexes was demonstrated for a range of membrane protein targets (Frauenfeld et al., 2016). The method is fast and requires few resources. The saposin–lipid–scaffold adapts to various sizes of transmembrane regions during self-assembly, forming a minimal lipid nanoparticle. This results in the formation of a well-defined membrane protein–lipid complex, which is desirable for structural characterization. Here, we describe a protocol to reconstitute the sarco-endoplasmic reticulum calcium ATPase (SERCA) into Salipro nanoparticles. The complex formation is analyzed using negative stain electron microscopy (EM), allowing to quickly determine an initial structure of the membrane protein and to evaluate sample conditions for structural studies using single-particle cryo-EM in a detergent-free environment.

AB - Membrane proteins depend on their natural lipid environment for function, which makes them more difficult to study in isolation. A number of approaches that mimic the lipid bilayer of biological membranes have been described (nanodiscs, SMALPs), enabling novel ways to assay activity and elucidate structures of this important class of proteins. More recently, the use of saposin A, a protein that is involved in lipid transport, to form Salipro (saposin–lipid–protein) complexes was demonstrated for a range of membrane protein targets (Frauenfeld et al., 2016). The method is fast and requires few resources. The saposin–lipid–scaffold adapts to various sizes of transmembrane regions during self-assembly, forming a minimal lipid nanoparticle. This results in the formation of a well-defined membrane protein–lipid complex, which is desirable for structural characterization. Here, we describe a protocol to reconstitute the sarco-endoplasmic reticulum calcium ATPase (SERCA) into Salipro nanoparticles. The complex formation is analyzed using negative stain electron microscopy (EM), allowing to quickly determine an initial structure of the membrane protein and to evaluate sample conditions for structural studies using single-particle cryo-EM in a detergent-free environment.

KW - Cryo-EM

KW - Detergent

KW - Detergent-free

KW - Electron microscopy

KW - Lipids

KW - Membrane protein structure

KW - P-type ATPases

KW - Salipro

KW - Saposin

KW - SERCA

UR - http://www.scopus.com/inward/record.url?scp=85024932823&partnerID=8YFLogxK

U2 - 10.1016/bs.mie.2017.06.035

DO - 10.1016/bs.mie.2017.06.035

M3 - Book chapter

C2 - 28779844

AN - SCOPUS:85024932823

SN - 978-0-12-812353-9

VL - 594

T3 - Methods in Enzymology

SP - 85

EP - 99

BT - Methods in Enzymology

A2 - Ziegler, Christine

PB - Academic Press

ER -