Molecular snapshots of dynamic membrane-bound metabolons

Jean Etienne Bassard*, Tomas Laursen

*Corresponding author af dette arbejde

Abstract

Numerous biosynthetic pathways have been shown to assemble at the surface of cellular membranes into efficient dynamic supramolecular assemblies termed metabolons. In response to environmental stimuli, metabolons assemble on-demand making them highly dynamic and fragile. This transient nature has previously hampered isolation and molecular characterization of dynamic metabolons. In contrast to conventional detergents, which tend to disrupt weak protein–protein interactions and remove lipids, the competence of a styrene maleic acid copolymer to carve out discrete lipid nanodisc from membranes offers immense potential for isolation of intact protein assemblies. Here, we present a method to extract the entire membrane-bound dhurrin pathway directly from microsomal fractions of the cereal Sorghum bicolor. This detergent-free nanodisc approach may be generally transposed for isolation of entire plant biosynthetic metabolons. This method provides a simple practical toolkit for the study of membrane protein complexes.

OriginalsprogEngelsk
TitelMethods in Enzymology
Antal sider27
Vol/bind617
ForlagAcademic Press
Publikationsdato1 jan. 2019
Sider1-27
Kapitel1
ISBN (Elektronisk)978-0-12-817074-8
DOI
StatusUdgivet - 1 jan. 2019
NavnMethods in Enzymology
Vol/bind617
ISSN0076-6879

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