A palette of fluorescent proteins optimized for diverse cellular environments

Lindsey M. Costantini, Mikhail Baloban, Michele L. Markwardt, Mark Rizzo, Feng Guo, Vladislav Verkhusha, Erik L. Snapp

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

To perform quantitative live cell imaging, investigators require fluorescent reporters that accurately report protein localization and levels, while minimally perturbing the cell. Yet, within the biochemically distinct environments of cellular organelles, popular fluorescent proteins (FPs), including EGFP, can be unreliable for quantitative imaging, resulting in the underestimation of protein levels and incorrect localization. Specifically, within the secretory pathway, significant populations of FPs misfold and fail to fluoresce due to non-native disulphide bond formation. Furthermore, transmembrane FP-fusion constructs can disrupt organelle architecture due to oligomerizing tendencies of numerous common FPs. Here, we describe a powerful set of bright and inert FPs optimized for use in multiple cellular compartments, especially oxidizing environments and biological membranes. Also, we provide new insights into the use of red FPs in the secretory pathway. Our monomeric 'oxFPs' finally resolve long-standing, underappreciated and important problems of cell biology and should be useful for a number of applications.

Original languageEnglish (US)
Article number8670
JournalNature Communications
Volume6
DOIs
StatePublished - Jul 9 2015

Fingerprint

proteins
Proteins
Secretory Pathway
Organelles
organelles
Cytology
Biological membranes
Imaging techniques
Disulfides
Cell Biology
disulfides
compartments
cells
Research Personnel
Fusion reactions
tendencies
Membranes
fusion
membranes
Population

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemistry(all)
  • Physics and Astronomy(all)

Cite this

Costantini, L. M., Baloban, M., Markwardt, M. L., Rizzo, M., Guo, F., Verkhusha, V., & Snapp, E. L. (2015). A palette of fluorescent proteins optimized for diverse cellular environments. Nature Communications, 6, [8670]. https://doi.org/10.1038/ncomms8670

A palette of fluorescent proteins optimized for diverse cellular environments. / Costantini, Lindsey M.; Baloban, Mikhail; Markwardt, Michele L.; Rizzo, Mark; Guo, Feng; Verkhusha, Vladislav; Snapp, Erik L.

In: Nature Communications, Vol. 6, 8670, 09.07.2015.

Research output: Contribution to journalArticle

Costantini, LM, Baloban, M, Markwardt, ML, Rizzo, M, Guo, F, Verkhusha, V & Snapp, EL 2015, 'A palette of fluorescent proteins optimized for diverse cellular environments', Nature Communications, vol. 6, 8670. https://doi.org/10.1038/ncomms8670
Costantini, Lindsey M. ; Baloban, Mikhail ; Markwardt, Michele L. ; Rizzo, Mark ; Guo, Feng ; Verkhusha, Vladislav ; Snapp, Erik L. / A palette of fluorescent proteins optimized for diverse cellular environments. In: Nature Communications. 2015 ; Vol. 6.
@article{a1218ee73d82401bb6ad1df5eeaf470a,
title = "A palette of fluorescent proteins optimized for diverse cellular environments",
abstract = "To perform quantitative live cell imaging, investigators require fluorescent reporters that accurately report protein localization and levels, while minimally perturbing the cell. Yet, within the biochemically distinct environments of cellular organelles, popular fluorescent proteins (FPs), including EGFP, can be unreliable for quantitative imaging, resulting in the underestimation of protein levels and incorrect localization. Specifically, within the secretory pathway, significant populations of FPs misfold and fail to fluoresce due to non-native disulphide bond formation. Furthermore, transmembrane FP-fusion constructs can disrupt organelle architecture due to oligomerizing tendencies of numerous common FPs. Here, we describe a powerful set of bright and inert FPs optimized for use in multiple cellular compartments, especially oxidizing environments and biological membranes. Also, we provide new insights into the use of red FPs in the secretory pathway. Our monomeric 'oxFPs' finally resolve long-standing, underappreciated and important problems of cell biology and should be useful for a number of applications.",
author = "Costantini, {Lindsey M.} and Mikhail Baloban and Markwardt, {Michele L.} and Mark Rizzo and Feng Guo and Vladislav Verkhusha and Snapp, {Erik L.}",
year = "2015",
month = "7",
day = "9",
doi = "10.1038/ncomms8670",
language = "English (US)",
volume = "6",
journal = "Nature Communications",
issn = "2041-1723",
publisher = "Nature Publishing Group",

}

TY - JOUR

T1 - A palette of fluorescent proteins optimized for diverse cellular environments

AU - Costantini, Lindsey M.

AU - Baloban, Mikhail

AU - Markwardt, Michele L.

AU - Rizzo, Mark

AU - Guo, Feng

AU - Verkhusha, Vladislav

AU - Snapp, Erik L.

PY - 2015/7/9

Y1 - 2015/7/9

N2 - To perform quantitative live cell imaging, investigators require fluorescent reporters that accurately report protein localization and levels, while minimally perturbing the cell. Yet, within the biochemically distinct environments of cellular organelles, popular fluorescent proteins (FPs), including EGFP, can be unreliable for quantitative imaging, resulting in the underestimation of protein levels and incorrect localization. Specifically, within the secretory pathway, significant populations of FPs misfold and fail to fluoresce due to non-native disulphide bond formation. Furthermore, transmembrane FP-fusion constructs can disrupt organelle architecture due to oligomerizing tendencies of numerous common FPs. Here, we describe a powerful set of bright and inert FPs optimized for use in multiple cellular compartments, especially oxidizing environments and biological membranes. Also, we provide new insights into the use of red FPs in the secretory pathway. Our monomeric 'oxFPs' finally resolve long-standing, underappreciated and important problems of cell biology and should be useful for a number of applications.

AB - To perform quantitative live cell imaging, investigators require fluorescent reporters that accurately report protein localization and levels, while minimally perturbing the cell. Yet, within the biochemically distinct environments of cellular organelles, popular fluorescent proteins (FPs), including EGFP, can be unreliable for quantitative imaging, resulting in the underestimation of protein levels and incorrect localization. Specifically, within the secretory pathway, significant populations of FPs misfold and fail to fluoresce due to non-native disulphide bond formation. Furthermore, transmembrane FP-fusion constructs can disrupt organelle architecture due to oligomerizing tendencies of numerous common FPs. Here, we describe a powerful set of bright and inert FPs optimized for use in multiple cellular compartments, especially oxidizing environments and biological membranes. Also, we provide new insights into the use of red FPs in the secretory pathway. Our monomeric 'oxFPs' finally resolve long-standing, underappreciated and important problems of cell biology and should be useful for a number of applications.

UR - http://www.scopus.com/inward/record.url?scp=84935506373&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84935506373&partnerID=8YFLogxK

U2 - 10.1038/ncomms8670

DO - 10.1038/ncomms8670

M3 - Article

VL - 6

JO - Nature Communications

JF - Nature Communications

SN - 2041-1723

M1 - 8670

ER -