Aarhus University Seal / Aarhus Universitets segl

Claus Elsborg Olesen

Lipid exchange by ultracentrifugation

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

Lipids play an important role in maintaining P-type ATPase structure and function, and often they are crucial for ATPase activity. When the P-type ATPases are in the membrane, they are surrounded by a mix of different lipids species with varying aliphatic chain lengths and saturation, and the complex interplay between the lipids and the P-type ATPases are still not well understood. We here describe a robust method to exchange the majority of the lipids surrounding the ATPase after solubilisation and/or purification with a target lipid of interest. The method is based on an ultracentrifugation step, where the protein sample is spun through a dense buffer containing large excess of the target lipid, which results in an approximately 80-85 % lipid exchange. The method is a very gently technique that maintains protein folding during the process, hence allowing further characterization of the protein in the presence of a target lipid of interest.
OriginalsprogEngelsk
TitelP-Type ATPases. Methods and Protocols
UdgivelsesstedNew York
ForlagSpringer
Udgivelsesår2016
Sider397-402
ISBN (trykt)978-1-4939-3178-1
StatusUdgivet - 2016
SerietitelMethods in Molecular Biology
Vol/bind1377
ISSN1064-3745

    Forskningsområder

  • Native lipids, Synthetic lipids, lipid exchange, Ultracentrifugation, density gradient, Membrane proteins, SERCA

Se relationer på Aarhus Universitet Citationsformater

ID: 85257791