English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Photophysical Behavior of mNeonGreen, an Evolutionarily Distant Green Fluorescent Protein

Steiert, F., Petrov, E. P., Schultz, P., Schwille, P., & Weidemann, T. (2018). Photophysical Behavior of mNeonGreen, an Evolutionarily Distant Green Fluorescent Protein. Biophysical Journal, 114(10), 2419-2431. doi:10.1016/j.bpj.2018.04.013.

Item is

Basic

show hide
Genre: Journal Article

Files

show Files
hide Files
:
1-s2.0-S0006349518304545-main.pdf (Publisher version), 989KB
Name:
1-s2.0-S0006349518304545-main.pdf
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
open archive

Locators

show

Creators

show
hide
 Creators:
Steiert, Frederik1, Author              
Petrov, Eugene P.1, Author              
Schultz, Peter1, Author              
Schwille, Petra1, Author              
Weidemann, Thomas1, Author              
Affiliations:
1Schwille, Petra / Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565169              

Content

show
hide
Free keywords: NONNEGATIVE MATRIX FACTORIZATION; CORRELATION SPECTROSCOPY; STRUCTURAL BASIS; BRANCHIOSTOMA-LANCEOLATUM; CRYSTAL-STRUCTURE; PROTON-TRANSFER; IN-VIVO; DYNAMICS; CHROMOPHORE; BRIGHTBiophysics;
 Abstract: Fluorescent proteins (FPs) feature complex photophysical behavior that must be considered when studying the dynamics of fusion proteins in model systems and live cells. In this work, we characterize mNeonGreen (mNG), a recently introduced FP from the bilaterian Branchiostoma lanceolatum, in comparison to the well-known hydrozoan variants enhanced green fluorescent protein (EGFP) and Aequorea coerulescens GFP by steady-state spectroscopy and fluorescence correlation spectroscopy in solutions of different pH. Blind spectral unmixing of sets of absorption spectra reveals three interconverting electronic states of mNG: a nonfluorescent protonated state, a bright state showing bell-shaped pH dependence, and a similarly bright state dominating at high pH. The gradual population of the acidic form by external protonation is reflected by increased flickering at low pH in fluorescence correlation spectroscopy measurements, albeit with much slower flicker rates and lower amplitudes as compared to Aequorea GFPs. In addition, increased flickering of mNG indicates a second deprotonation step above pH 10 leading to a slight decrease in fluorescence. Thus, mNG is distinguished from Aequorea GFPs by a two-step protonation response with opposite effects that reflects a chemically distinct chromophore environment. Despite the more complex pH dependence, mNG represents a superior FP under a broad range of conditions.

Details

show
hide
Language(s): eng - English
 Dates: 2018
 Publication Status: Published in print
 Pages: 13
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: ISI: 000432700300019
DOI: 10.1016/j.bpj.2018.04.013
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Biophysical Journal
  Other : Biophys. J.
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: Cambridge, Mass. : Cell Press
Pages: - Volume / Issue: 114 (10) Sequence Number: - Start / End Page: 2419 - 2431 Identifier: ISSN: 0006-3495
CoNE: https://pure.mpg.de/cone/journals/resource/954925385117