title: Single-Molecule Specific Mislocalization of Red Fluorescent Proteins in Live Escherichia coli.
abstract: Tagging of individual proteins with genetically encoded fluorescent proteins (FPs) has been used extensively to study localization and interactions in live cells. Recent developments in single-molecule localization microscopy have enabled the dynamic visualization of individual tagged proteins inside living cells. However, tagging proteins with FPs is not without problems: formation of insoluble aggregates and inhibition of native functions of the protein are well-known issues. Previously reported artifacts manifest themselves at all expression levels of the FP-tagged proteins, making the design of control experiments relatively straightforward. Here, we describe a previously uncharacterized mislocalization artifact of Entacmaea quadricolor red fluorescent protein variants that is detectable at the single-molecule level in live Escherichia coli cells.
authors.affiliation: Zernike Institute for Advanced Materials, Rijksuniversiteit Groningen, Groningen, The Netherlands; School of Chemistry, University of Wollongong, Wollongong, New South Wales, Australia.
authors.forename: Harshad
authors.initials: H
authors.lastname: Ghodke
authors.affiliation: Zernike Institute for Advanced Materials, Rijksuniversiteit Groningen, Groningen, The Netherlands.
authors.forename: Victor E A
authors.initials: VE
authors.lastname: Caldas
authors.affiliation: Zernike Institute for Advanced Materials, Rijksuniversiteit Groningen, Groningen, The Netherlands.
authors.forename: Christiaan M
authors.initials: CM
authors.lastname: Punter
authors.affiliation: School of Chemistry, University of Wollongong, Wollongong, New South Wales, Australia. Electronic address: vanoijen@uow.edu.au.
authors.forename: Antoine M
authors.initials: AM
authors.lastname: van Oijen
authors.affiliation: Zernike Institute for Advanced Materials, Rijksuniversiteit Groningen, Groningen, The Netherlands; School of Chemistry, University of Wollongong, Wollongong, New South Wales, Australia.
authors.forename: Andrew
authors.initials: A
authors.lastname: Robinson
mesh_terms: D004926:Escherichia coli / Q000378:metabolism* / Q000502:physiology; D008164:Luminescent Proteins / Q000378:metabolism*; D050296:Microbial Viability; D021381:Protein Transport; D000072760:Single Molecule Imaging; D000097573:Red Fluorescent Protein
chemical_list: D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein