Near-Infrared Fluorescent Proteins Engineered from Bacterial Phytochromes in Neuroimaging

Kiryl D. Piatkevich, Ho Jun Suk, Suhasa B. Kodandaramaiah, Fumiaki Yoshida, Ellen M. DeGennaro, Mikhail Drobizhev, Thomas E. Hughes, Robert Desimone, Edward S. Boyden, Vladislav Verkhusha

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Several series of near-infrared (NIR) fluorescent proteins (FPs) were recently engineered from bacterial phytochromes but were not systematically compared in neurons. To fluoresce, NIR FPs utilize an enzymatic derivative of heme, the linear tetrapyrrole biliverdin, as a chromophore whose level in neurons is poorly studied. Here, we evaluated NIR FPs of the iRFP protein family, which were reported to be the brightest in non-neuronal mammalian cells, in primary neuronal culture, in brain slices of mouse and monkey, and in mouse brain in vivo. We applied several fluorescence imaging modes, such as wide-field and confocal one-photon and two-photon microscopy, to compare photochemical and biophysical properties of various iRFPs. The iRFP682 and iRFP670 proteins exhibited the highest brightness and photostability under one-photon and two-photon excitation modes, respectively. All studied iRFPs exhibited efficient binding of the endogenous biliverdin chromophore in cultured neurons and in the mammalian brain and can be readily applied to neuroimaging.

Original languageEnglish (US)
JournalBiophysical Journal
DOIs
StateAccepted/In press - 2017

Fingerprint

Phytochrome
Bacterial Proteins
Neuroimaging
Photons
Biliverdine
Proteins
Neurons
Brain
Tetrapyrroles
Optical Imaging
Heme
Haplorhini
Microscopy

ASJC Scopus subject areas

  • Biophysics

Cite this

Piatkevich, K. D., Suk, H. J., Kodandaramaiah, S. B., Yoshida, F., DeGennaro, E. M., Drobizhev, M., ... Verkhusha, V. (Accepted/In press). Near-Infrared Fluorescent Proteins Engineered from Bacterial Phytochromes in Neuroimaging. Biophysical Journal. https://doi.org/10.1016/j.bpj.2017.09.007

Near-Infrared Fluorescent Proteins Engineered from Bacterial Phytochromes in Neuroimaging. / Piatkevich, Kiryl D.; Suk, Ho Jun; Kodandaramaiah, Suhasa B.; Yoshida, Fumiaki; DeGennaro, Ellen M.; Drobizhev, Mikhail; Hughes, Thomas E.; Desimone, Robert; Boyden, Edward S.; Verkhusha, Vladislav.

In: Biophysical Journal, 2017.

Research output: Contribution to journalArticle

Piatkevich, KD, Suk, HJ, Kodandaramaiah, SB, Yoshida, F, DeGennaro, EM, Drobizhev, M, Hughes, TE, Desimone, R, Boyden, ES & Verkhusha, V 2017, 'Near-Infrared Fluorescent Proteins Engineered from Bacterial Phytochromes in Neuroimaging', Biophysical Journal. https://doi.org/10.1016/j.bpj.2017.09.007
Piatkevich, Kiryl D. ; Suk, Ho Jun ; Kodandaramaiah, Suhasa B. ; Yoshida, Fumiaki ; DeGennaro, Ellen M. ; Drobizhev, Mikhail ; Hughes, Thomas E. ; Desimone, Robert ; Boyden, Edward S. ; Verkhusha, Vladislav. / Near-Infrared Fluorescent Proteins Engineered from Bacterial Phytochromes in Neuroimaging. In: Biophysical Journal. 2017.
@article{661c960718b24c0d979bfb8a73c876a9,
title = "Near-Infrared Fluorescent Proteins Engineered from Bacterial Phytochromes in Neuroimaging",
abstract = "Several series of near-infrared (NIR) fluorescent proteins (FPs) were recently engineered from bacterial phytochromes but were not systematically compared in neurons. To fluoresce, NIR FPs utilize an enzymatic derivative of heme, the linear tetrapyrrole biliverdin, as a chromophore whose level in neurons is poorly studied. Here, we evaluated NIR FPs of the iRFP protein family, which were reported to be the brightest in non-neuronal mammalian cells, in primary neuronal culture, in brain slices of mouse and monkey, and in mouse brain in vivo. We applied several fluorescence imaging modes, such as wide-field and confocal one-photon and two-photon microscopy, to compare photochemical and biophysical properties of various iRFPs. The iRFP682 and iRFP670 proteins exhibited the highest brightness and photostability under one-photon and two-photon excitation modes, respectively. All studied iRFPs exhibited efficient binding of the endogenous biliverdin chromophore in cultured neurons and in the mammalian brain and can be readily applied to neuroimaging.",
author = "Piatkevich, {Kiryl D.} and Suk, {Ho Jun} and Kodandaramaiah, {Suhasa B.} and Fumiaki Yoshida and DeGennaro, {Ellen M.} and Mikhail Drobizhev and Hughes, {Thomas E.} and Robert Desimone and Boyden, {Edward S.} and Vladislav Verkhusha",
year = "2017",
doi = "10.1016/j.bpj.2017.09.007",
language = "English (US)",
journal = "Biophysical Journal",
issn = "0006-3495",
publisher = "Biophysical Society",

}

TY - JOUR

T1 - Near-Infrared Fluorescent Proteins Engineered from Bacterial Phytochromes in Neuroimaging

AU - Piatkevich, Kiryl D.

AU - Suk, Ho Jun

AU - Kodandaramaiah, Suhasa B.

AU - Yoshida, Fumiaki

AU - DeGennaro, Ellen M.

AU - Drobizhev, Mikhail

AU - Hughes, Thomas E.

AU - Desimone, Robert

AU - Boyden, Edward S.

AU - Verkhusha, Vladislav

PY - 2017

Y1 - 2017

N2 - Several series of near-infrared (NIR) fluorescent proteins (FPs) were recently engineered from bacterial phytochromes but were not systematically compared in neurons. To fluoresce, NIR FPs utilize an enzymatic derivative of heme, the linear tetrapyrrole biliverdin, as a chromophore whose level in neurons is poorly studied. Here, we evaluated NIR FPs of the iRFP protein family, which were reported to be the brightest in non-neuronal mammalian cells, in primary neuronal culture, in brain slices of mouse and monkey, and in mouse brain in vivo. We applied several fluorescence imaging modes, such as wide-field and confocal one-photon and two-photon microscopy, to compare photochemical and biophysical properties of various iRFPs. The iRFP682 and iRFP670 proteins exhibited the highest brightness and photostability under one-photon and two-photon excitation modes, respectively. All studied iRFPs exhibited efficient binding of the endogenous biliverdin chromophore in cultured neurons and in the mammalian brain and can be readily applied to neuroimaging.

AB - Several series of near-infrared (NIR) fluorescent proteins (FPs) were recently engineered from bacterial phytochromes but were not systematically compared in neurons. To fluoresce, NIR FPs utilize an enzymatic derivative of heme, the linear tetrapyrrole biliverdin, as a chromophore whose level in neurons is poorly studied. Here, we evaluated NIR FPs of the iRFP protein family, which were reported to be the brightest in non-neuronal mammalian cells, in primary neuronal culture, in brain slices of mouse and monkey, and in mouse brain in vivo. We applied several fluorescence imaging modes, such as wide-field and confocal one-photon and two-photon microscopy, to compare photochemical and biophysical properties of various iRFPs. The iRFP682 and iRFP670 proteins exhibited the highest brightness and photostability under one-photon and two-photon excitation modes, respectively. All studied iRFPs exhibited efficient binding of the endogenous biliverdin chromophore in cultured neurons and in the mammalian brain and can be readily applied to neuroimaging.

UR - http://www.scopus.com/inward/record.url?scp=85030651185&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85030651185&partnerID=8YFLogxK

U2 - 10.1016/j.bpj.2017.09.007

DO - 10.1016/j.bpj.2017.09.007

M3 - Article

C2 - 29017728

AN - SCOPUS:85030651185

JO - Biophysical Journal

JF - Biophysical Journal

SN - 0006-3495

ER -