Nanodisc self-assembly is thermodynamically reversible and controllable

Tyler Camp, Stephen G. Sligar, Stephen G. Sligar, Stephen G. Sligar

Research output: Contribution to journalArticlepeer-review

Abstract

Many highly ordered complex systems form by the spontaneous self-assembly of simpler subunits. An important biophysical tool that relies on self-assembly is the Nanodisc system, which finds extensive use as native-like environments for studying membrane proteins. Nanodiscs are self-assembled from detergent-solubilized mixtures of phospholipids and engineered helical proteins called membrane scaffold proteins (MSPs). Detergent removal results in the formation of nanoscale bilayers stabilized by two MSP "belts."Despite their numerous applications in biology, and contributions from many laboratories world-wide, little is known about the self-assembly process such as when the bilayer forms or when the MSP associates with lipids. We use fluorescence and optical spectroscopy to probe self-assembly at various equilibria defined by the detergent concentration. We show that the bilayer begins forming below the critical micellar concentration of the detergent (10 mM), and the association of MSP and lipids begins at lower detergent levels, showing a dependence on the concentrations of MSP and lipids. Following the dissolution process by adding detergent to purified Nanodiscs demonstrates that the self-assembly is reversible. Our data demonstrate that Nanodisc self-assembly is experimentally accessible, and that controlling the detergent concentration allows exquisite control over the self-assembly reaction. This improved understanding of self-assembly could lead to better functional incorporation of hitherto intractable membrane target proteins. This journal is

Original languageEnglish (US)
Pages (from-to)5615-5623
Number of pages9
JournalSoft Matter
Volume16
Issue number24
DOIs
StatePublished - Jun 28 2020

ASJC Scopus subject areas

  • Chemistry(all)
  • Condensed Matter Physics

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