title: Sortase-mediated labelling of lipid nanodiscs for cellular tracing.
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.
authors.affiliation: School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, UK. m.e.webb@leeds.ac.uk p.a.beales@leeds.ac.uk.
authors.forename: A Ivona
authors.initials: AI
authors.lastname: Petrache
authors.affiliation: School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, UK. m.e.webb@leeds.ac.uk p.a.beales@leeds.ac.uk.
authors.forename: Darren C
authors.initials: DC
authors.lastname: Machin
authors.affiliation: School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, UK. m.e.webb@leeds.ac.uk p.a.beales@leeds.ac.uk.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Williamson
authors.affiliation: School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, UK. m.e.webb@leeds.ac.uk p.a.beales@leeds.ac.uk.
authors.forename: Michael E
authors.initials: ME
authors.lastname: Webb
authors.affiliation: School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, UK. m.e.webb@leeds.ac.uk p.a.beales@leeds.ac.uk.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Beales
mesh_terms: D019881:Aminoacyltransferases / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D058948:Cell Tracking / Q000379:methods; D003546:Cysteine Endopeptidases / Q000737:chemistry*; D000067493:Dynamic Light Scattering; D006367:HeLa Cells; D006801:Humans; D008055:Lipids / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D008856:Microscopy, Fluorescence; D049329:Nanostructures / Q000737:chemistry*; D010316:Particle Size; D013194:Staining and Labeling
chemical_list: D001426:Bacterial Proteins; D008055:Lipids; D008565:Membrane Proteins; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases