[NMR paper] A triplet state mechanism of primed conversion enables rational engineering of photoconvertible fluorescent proteins for dual color fluorescence nanoscopy
A triplet state mechanism of primed conversion enables rational engineering of photoconvertible fluorescent proteins for dual color fluorescence nanoscopy
Photoconvertible fluorescent proteins (pcFPs) are powerful tools for super-resolution localization microscopy and protein tagging. Recently, they have been found to undergo efficient photoconversion by an alternative method termed primed conversion, employing dual wavelength illumination with blue and far-red/near-infrared light. Primed conversion has been reported only for Dendra2 and its mechanism has remained elusive. Here, we uncover the molecular mechanism of primed conversion by reporting the intermediate "primed" state to be a triplet dark state formed by intersystem crossing. We show that formation of this state can be influenced by the introduction of serine or threonine at sequence position 69 (Eos notation) and use this knowledge to create "pr"- (for primed convertible) variants of most known green-to-red pcFPs. Among these, pr-mEos2 exhibits excellent properties for primed PALM, allowing a new dual color PALM modality that does not occupy additional spectral space.
[NMR paper] A general mechanism of photoconversion of green-to-red fluorescent proteins based on blue and infrared light reduces phototoxicity in live-cell single-molecule imaging
A general mechanism of photoconversion of green-to-red fluorescent proteins based on blue and infrared light reduces phototoxicity in live-cell single-molecule imaging
Photoconversion of fluorescent proteins by blue and complementary near-infrared light, termed primed conversion (PC), is a mechanism recently discovered for Dendra2. We demonstrate that controlling the conformation of arginine at residue 66 by threonine at residue 69 of fluorescent proteins from Anthozoan families (Dendra2, mMaple, Eos, mKikGR, pcDronpa protein families) represents a general route to facilitate PC....
[NMR paper] Bioconjugation of proteins with a paramagnetic NMR and fluorescent tag.
Bioconjugation of proteins with a paramagnetic NMR and fluorescent tag.
Related Articles Bioconjugation of proteins with a paramagnetic NMR and fluorescent tag.
Chemistry. 2013 Dec 9;19(50):17141-9
Authors: Huang F, Pei YY, Zuo HH, Chen JL, Yang Y, Su XC
Abstract
Site-specific labeling of proteins with lanthanide ions offers great opportunities for investigating the structure, function, and dynamics of proteins by virtue of the unique properties of lanthanides. Lanthanide-tagged proteins can be studied by NMR, X-ray, fluorescence, and EPR...
nmrlearner
Journal club
0
12-07-2013 01:00 PM
Matrix-free dynamic nuclear polarization enables solid-state NMR (13)C-(13)C correlation spectroscopy of proteins at natural isotopic abundance
From The DNP-NMR Blog:
Matrix-free dynamic nuclear polarization enables solid-state NMR (13)C-(13)C correlation spectroscopy of proteins at natural isotopic abundance
Takahashi, H., S. Hediger, and G. De Paepe, Matrix-free dynamic nuclear polarization enables solid-state NMR (13)C-(13)C correlation spectroscopy of proteins at natural isotopic abundance. Chem Commun (Camb), 2013. 49(82): p. 9479-81.
http://www.ncbi.nlm.nih.gov/pubmed/24013616
nmrlearner
News from NMR blogs
0
11-21-2013 01:14 AM
[NMR paper] Matrix-free dynamic nuclear polarization enables solid-state NMR (13)C-(13)C correlation spectroscopy of proteins at natural isotopic abundance.
Matrix-free dynamic nuclear polarization enables solid-state NMR (13)C-(13)C correlation spectroscopy of proteins at natural isotopic abundance.
Matrix-free dynamic nuclear polarization enables solid-state NMR (13)C-(13)C correlation spectroscopy of proteins at natural isotopic abundance.
Chem Commun (Camb). 2013 Sep 6;
Authors: Takahashi H, Hediger S, De Paëpe G
Abstract
We introduce a general approach for dynamic nuclear polarization (DNP) enhanced solid-state NMR that overcomes the current problems in DNP experiments caused by the use...
nmrlearner
Journal club
0
09-10-2013 08:44 PM
[NMR900 blog] Solid-State NMR in Materials for Energy Storage and Conversion
Solid-State NMR in Materials for Energy Storage and Conversion
A special issue of Solid State Nuclear Magnetic Resonance guest-edited by Clare Grey (Cambridge) and Gillian Goward (McMaster), Volume 42, Pages 1-98 (April 2012) http://www.sciencedirect.com/science/journal/09262040/42
Editorial
Clare P. Grey, Gillian R. Goward, Editorial "Solid-State NMR in Materials for Energy Storage and Conversion," Solid State Nuclear Magnetic Resonance
42 (2012) 1. http://dx.doi.org/10.1016/j.ssnmr.2012.03.001
Canadian contributions
nmrlearner
News from NMR blogs
0
03-21-2012 11:40 PM
Systematic Study of Protein Detection Mechanism of Self-Assembling 19F NMR/MRI Nanoprobes toward Rational Design and Improved Sensitivity
Systematic Study of Protein Detection Mechanism of Self-Assembling 19F NMR/MRI Nanoprobes toward Rational Design and Improved Sensitivity
Yousuke Takaoka, Keishi Kiminami, Keigo Mizusawa, Kazuya Matsuo, Michiko Narazaki, Tetsuya Matsuda and Itaru Hamachi
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/ja203996c/aop/images/medium/ja-2011-03996c_0004.gif
Journal of the American Chemical Society
DOI: 10.1021/ja203996c
http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/jacsat/~4/fqTSjFalrGg
A triplet state mechanism of primed conversion enables rational engineering of photoconvertible fluorescent proteins for dual color fluorescence nanoscopy
Linear Mode
