Confocal Single-Molecule FRET for Protein Conformational Dynamics

2013 ◽  
pp. 51-62 ◽  
Author(s):  
Yan-Wen Tan ◽  
Jeffrey A. Hanson ◽  
Jhih-Wei Chu ◽  
Haw Yang
Keyword(s):  
Single Molecule ◽  
Conformational Dynamics ◽  
Single Molecule Fret ◽  
Protein Conformational Dynamics

scholarly journals Protein Conformational Dynamics In Cellula and IN VIVO Using Live-Cell Single-Molecule Fret Hilo Microscopy

2021 ◽  
Vol 120 (3) ◽  
pp. 114a
Author(s):  
Abhinaya Anandamurugan ◽  
Philipp Wortmann ◽  
Chenyang Lan ◽  
April Garcia Fernando ◽  
Marina Veil ◽  
...  
Keyword(s):  
Single Molecule ◽  
Conformational Dynamics ◽  
Live Cell ◽  
Single Molecule Fret ◽  
Protein Conformational Dynamics

Conformational Dynamics of DNA Hairpins at Millisecond Resolution Obtained from Analysis of Single-Molecule FRET Histograms

2013 ◽  
Vol 117 (50) ◽  
pp. 16105-16109 ◽  
Author(s):  
Roman Tsukanov ◽  
Toma E. Tomov ◽  
Yaron Berger ◽  
Miran Liber ◽  
Eyal Nir
Keyword(s):  
Single Molecule ◽  
Conformational Dynamics ◽  
Dna Hairpins ◽  
Single Molecule Fret

scholarly journals Conformational dynamics of the ABC transporter McjD seen by single‐molecule FRET

2018 ◽  
Vol 37 (21) ◽  
Author(s):  
Florence Husada ◽  
Kiran Bountra ◽  
Konstantinos Tassis ◽  
Marijn Boer ◽  
Maria Romano ◽  
...  
Keyword(s):  
Abc Transporter ◽  
Single Molecule ◽  
Conformational Dynamics ◽  
Single Molecule Fret

scholarly journals Single Molecule FRET Studies of Protein Conformational Landscapes: 3 Prototypic Examples for the Relation Between Conformational Dynamics and Function

2011 ◽  
Vol 100 (3) ◽  
pp. 474a-475a
Author(s):  
Markus Richert ◽  
Dymitro Rodnin ◽  
Carola S. Hengstenberg ◽  
Thomas Peulen ◽  
Alessandro Valeri ◽  
...  
Keyword(s):  
Single Molecule ◽  
Conformational Dynamics ◽  
Single Molecule Fret ◽  
Dynamics And Function ◽  
And Function

scholarly journals 631: Single-molecule FRET reveals CFTR conformational dynamics during channel gating

2021 ◽  
Vol 20 ◽  
pp. S299-S300
Author(s):  
P. Liyanage ◽  
K. Mun ◽  
S. Yarlagadda ◽  
Y. Huang ◽  
A. Naren
Keyword(s):  
Single Molecule ◽  
Conformational Dynamics ◽  
Channel Gating ◽  
Single Molecule Fret

scholarly journals Conformational dynamics of Cas9 governing DNA cleavage revealed by single molecule FRET

2017 ◽  
Author(s):  
Mengyi Yang ◽  
Sijia Peng ◽  
Ruirui Sun ◽  
Jingdi Lin ◽  
Nan Wang ◽  
...  
Keyword(s):  
Single Molecule ◽  
Dna Cleavage ◽  
Conformational Dynamics ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Nuclease Activity ◽  
Single Molecule Fluorescence ◽  
Single Molecule Fret ◽  
Allosteric Communication ◽  
Target Binding

SummaryOff-target binding and cleavage by Cas9 pose as major challenges in its applications. How conformational dynamics of Cas9 governs its nuclease activity under on- and off-target conditions remains largely unknown. Here, using intra-molecular single molecule fluorescence resonance energy transfer measurements, we revealed that Cas9 in apo, sgRNA-bound, and dsDNA/sgRNA-bound forms all spontaneously transits between three major conformational states, mainly reflecting significant conformational mobility of the catalytic HNH domain. We furthermore uncovered a surprising long-range allosteric communication between the HNH domain and RNA/DNA heteroduplex at the PAM-distal end to ensure correct positioning of the catalytic site, which demonstrated a unique proofreading mechanism served as the last checkpoint before DNA cleavage. Several Cas9 residues were likely to mediate the allosteric communication and proofreading step. Modulating interactions between Cas9 and heteroduplex at the distal end by introducing mutations on these sites provides an alternative route to improve and optimize the CRISPR/Cas9 toolbox.

scholarly journals Exploring the effects of cosolutes and crowding on the volumetric and kinetic profile of the conformational dynamics of a poly dA loop DNA hairpin: a single-molecule FRET study

2018 ◽  
Vol 47 (2) ◽  
pp. 981-996 ◽  
Author(s):  
Satyajit Patra ◽  
Vitor Schuabb ◽  
Irena Kiesel ◽  
Jim-Marcel Knop ◽  
Rosario Oliva ◽  
...  
Keyword(s):  
Single Molecule ◽  
Conformational Dynamics ◽  
Dna Hairpin ◽  
Single Molecule Fret ◽  
Kinetic Profile
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