Dark, Ultra-Dark and Ultra-Bright Nanodiscs for membrane protein investigations

Mark A. McLean, Ilia G. Denisov, Yelena V. Grinkova, Stephen G. Sligar

Research output: Contribution to journalArticlepeer-review

Abstract

We describe the construction, expression and purification of three new membrane scaffold proteins (MSP) for use in assembling Nanodiscs. These new MSPs have a variety of luminescent properties for use in combination with several analytical methods. “Dark” MSP has no tryptophan residues, “Ultra-Dark” replaces both tryptophan and tyrosine with non-fluorescent side chains, and “Ultra-Bright” adds additional tryptophans to the parent membrane scaffold protein to provide a dramatic increase in native tryptophan fluorescence. All MSPs were used to successfully assemble Nanodiscs nominally 10 nm in diameter, and the resultant bilayer structure was characterized. An example of the usefulness of these new scaffold proteins is provided.

Original languageEnglish (US)
Article number113860
JournalAnalytical Biochemistry
Volume607
DOIs
StatePublished - Oct 15 2020

Keywords

  • Bilayer binding
  • Membrane mimetic
  • Membrane protein
  • Membrane scaffold protein
  • Nanodiscs

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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