Sortase-mediated labelling of lipid nanodiscs for cellular tracing

Mol Biosyst. 2016 May 24;12(6):1760-3. doi: 10.1039/c6mb00126b.

Abstract

Lipid nanodiscs have broad applications in membrane protein assays, biotechnology and materials science. Chemical modification of the nanodiscs to expand their functional attributes is generally desirable for all of these uses. We present a method for site-selective labelling of the N-terminus of the nanodisc's membrane scaffold protein (MSP) using the Sortase A protein. Labelling of the MSP was achieved when assembled within the lipid nanodisc architecture, demonstrating that this method can be used as a retrofit approach to modification of preformed nanodiscs before or during application. We label the MSP with a fluorescent fluorescein moiety and use them to image nanodisc uptake into HeLa cells. The Sortase A labelling method could be employed as a general approach to labelling nanodiscs with application-specific functionalities.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aminoacyltransferases / chemistry*
  • Bacterial Proteins / chemistry*
  • Cell Tracking* / methods
  • Cysteine Endopeptidases / chemistry*
  • Dynamic Light Scattering
  • HeLa Cells
  • Humans
  • Lipids / chemistry*
  • Membrane Proteins / chemistry
  • Membrane Proteins / metabolism
  • Microscopy, Fluorescence
  • Nanostructures / chemistry*
  • Particle Size
  • Staining and Labeling*

Substances

  • Bacterial Proteins
  • Lipids
  • Membrane Proteins
  • Aminoacyltransferases
  • sortase A
  • Cysteine Endopeptidases