A photoswitchable orange-to-far-red fluorescent protein, PSmOrange

Oksana M. Subach, George H. Patterson, Li Min Ting, Yarong Wang, John S. Condeelis, Vladislav Verkhusha

Research output: Contribution to journalArticle

96 Citations (Scopus)

Abstract

We report a photoswitchable monomeric Orange (PSmOrange) protein that is initially orange (excitation, 548 nm; emission, 565 nm) but becomes far-red (excitation, 636 nm; emission, 662 nm) after irradiation with blue-green light. Compared to its parental orange proteins, PSmOrange has greater brightness, faster maturation, higher photoconversion contrast and better photostability. The red-shifted spectra of both forms of PSmOrange enable its simultaneous use with cyan-to-green photoswitchable proteins to study four intracellular populations. Photoconverted PSmOrange has, to our knowledge, the most far-red excitation peak of all GFP-like fluorescent proteins, provides diffraction-limited and super-resolution imaging in the far-red light range, is optimally excited with common red lasers, and can be photoconverted subcutaneously in a mouse. PSmOrange photoswitching occurs via a two-step photo-oxidation process, which causes cleavage of the polypeptide backbone. The far-red fluorescence of photoconverted PSmOrange results from a new chromophore containing N-acylimine with a co-planar carbon-oxygen double bond.

Original languageEnglish (US)
Pages (from-to)771-780
Number of pages10
JournalNature Methods
Volume8
Issue number9
DOIs
StatePublished - Sep 2011

Fingerprint

Proteins
Methyl Green
Photooxidation
Chromophores
Luminance
Carbon
Diffraction
Fluorescence
Irradiation
Oxygen
Imaging techniques
Peptides
red fluorescent protein
Lasers
Light
Population

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology
  • Biochemistry
  • Cell Biology

Cite this

A photoswitchable orange-to-far-red fluorescent protein, PSmOrange. / Subach, Oksana M.; Patterson, George H.; Ting, Li Min; Wang, Yarong; Condeelis, John S.; Verkhusha, Vladislav.

In: Nature Methods, Vol. 8, No. 9, 09.2011, p. 771-780.

Research output: Contribution to journalArticle

Subach, Oksana M. ; Patterson, George H. ; Ting, Li Min ; Wang, Yarong ; Condeelis, John S. ; Verkhusha, Vladislav. / A photoswitchable orange-to-far-red fluorescent protein, PSmOrange. In: Nature Methods. 2011 ; Vol. 8, No. 9. pp. 771-780.
@article{f1229f391caa42569aa2ff0543996f54,
title = "A photoswitchable orange-to-far-red fluorescent protein, PSmOrange",
abstract = "We report a photoswitchable monomeric Orange (PSmOrange) protein that is initially orange (excitation, 548 nm; emission, 565 nm) but becomes far-red (excitation, 636 nm; emission, 662 nm) after irradiation with blue-green light. Compared to its parental orange proteins, PSmOrange has greater brightness, faster maturation, higher photoconversion contrast and better photostability. The red-shifted spectra of both forms of PSmOrange enable its simultaneous use with cyan-to-green photoswitchable proteins to study four intracellular populations. Photoconverted PSmOrange has, to our knowledge, the most far-red excitation peak of all GFP-like fluorescent proteins, provides diffraction-limited and super-resolution imaging in the far-red light range, is optimally excited with common red lasers, and can be photoconverted subcutaneously in a mouse. PSmOrange photoswitching occurs via a two-step photo-oxidation process, which causes cleavage of the polypeptide backbone. The far-red fluorescence of photoconverted PSmOrange results from a new chromophore containing N-acylimine with a co-planar carbon-oxygen double bond.",
author = "Subach, {Oksana M.} and Patterson, {George H.} and Ting, {Li Min} and Yarong Wang and Condeelis, {John S.} and Vladislav Verkhusha",
year = "2011",
month = "9",
doi = "10.1038/nmeth.1664",
language = "English (US)",
volume = "8",
pages = "771--780",
journal = "Nature Methods",
issn = "1548-7091",
publisher = "Nature Publishing Group",
number = "9",

}

TY - JOUR

T1 - A photoswitchable orange-to-far-red fluorescent protein, PSmOrange

AU - Subach, Oksana M.

AU - Patterson, George H.

AU - Ting, Li Min

AU - Wang, Yarong

AU - Condeelis, John S.

AU - Verkhusha, Vladislav

PY - 2011/9

Y1 - 2011/9

N2 - We report a photoswitchable monomeric Orange (PSmOrange) protein that is initially orange (excitation, 548 nm; emission, 565 nm) but becomes far-red (excitation, 636 nm; emission, 662 nm) after irradiation with blue-green light. Compared to its parental orange proteins, PSmOrange has greater brightness, faster maturation, higher photoconversion contrast and better photostability. The red-shifted spectra of both forms of PSmOrange enable its simultaneous use with cyan-to-green photoswitchable proteins to study four intracellular populations. Photoconverted PSmOrange has, to our knowledge, the most far-red excitation peak of all GFP-like fluorescent proteins, provides diffraction-limited and super-resolution imaging in the far-red light range, is optimally excited with common red lasers, and can be photoconverted subcutaneously in a mouse. PSmOrange photoswitching occurs via a two-step photo-oxidation process, which causes cleavage of the polypeptide backbone. The far-red fluorescence of photoconverted PSmOrange results from a new chromophore containing N-acylimine with a co-planar carbon-oxygen double bond.

AB - We report a photoswitchable monomeric Orange (PSmOrange) protein that is initially orange (excitation, 548 nm; emission, 565 nm) but becomes far-red (excitation, 636 nm; emission, 662 nm) after irradiation with blue-green light. Compared to its parental orange proteins, PSmOrange has greater brightness, faster maturation, higher photoconversion contrast and better photostability. The red-shifted spectra of both forms of PSmOrange enable its simultaneous use with cyan-to-green photoswitchable proteins to study four intracellular populations. Photoconverted PSmOrange has, to our knowledge, the most far-red excitation peak of all GFP-like fluorescent proteins, provides diffraction-limited and super-resolution imaging in the far-red light range, is optimally excited with common red lasers, and can be photoconverted subcutaneously in a mouse. PSmOrange photoswitching occurs via a two-step photo-oxidation process, which causes cleavage of the polypeptide backbone. The far-red fluorescence of photoconverted PSmOrange results from a new chromophore containing N-acylimine with a co-planar carbon-oxygen double bond.

UR - http://www.scopus.com/inward/record.url?scp=80052281149&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80052281149&partnerID=8YFLogxK

U2 - 10.1038/nmeth.1664

DO - 10.1038/nmeth.1664

M3 - Article

C2 - 21804536

AN - SCOPUS:80052281149

VL - 8

SP - 771

EP - 780

JO - Nature Methods

JF - Nature Methods

SN - 1548-7091

IS - 9

ER -