Single-Molecule Förster Resonance Energy Transfer Measurement Reveals the Dynamic Partially Ordered Structure of the Epidermal Growth Factor Receptor C-Tail Domain

2018 ◽  
Vol 123 (3) ◽  
pp. 571-581 ◽  
Author(s):  
Kenji Okamoto ◽  
Yasushi Sako
Keyword(s):  
Epidermal Growth Factor Receptor ◽  
Energy Transfer ◽  
Single Molecule ◽  
Resonance Energy Transfer ◽  
Growth Factor Receptor ◽  
Resonance Energy ◽  
Ordered Structure ◽  
Tail Domain ◽  
Transfer Measurement ◽  
Epidermal Growth

scholarly journals Single-Molecular Förster Resonance Energy Transfer Measurement on Structures and Interactions of Biomolecules

Micromachines ◽  
2021 ◽  
Vol 12 (5) ◽  
pp. 492
Author(s):  
Yi Qiao ◽  
Yuhan Luo ◽  
Naiyun Long ◽  
Yi Xing ◽  
Jing Tu
Keyword(s):  
Energy Transfer ◽  
Single Molecule ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Forster Resonance Energy Transfer ◽  
Förster Resonance Energy Transfer ◽  
Widespread Application ◽  
Novel Studies ◽  
Transfer Measurement ◽  
Förster Resonance

Single-molecule Förster resonance energy transfer (smFRET) inherits the strategy of measurement from the effective “spectroscopic ruler” FRET and can be utilized to observe molecular behaviors with relatively high throughput at nanometer scale. The simplicity in principle and configuration of smFRET make it easy to apply and couple with other technologies to comprehensively understand single-molecule dynamics in various application scenarios. Despite its widespread application, smFRET is continuously developing and novel studies based on the advanced platforms have been done. Here, we summarize some representative examples of smFRET research of recent years to exhibit the versatility and note typical strategies to further improve the performance of smFRET measurement on different biomolecules.

scholarly journals Enhanced dimerization drives ligand-independent activity of mutant epidermal growth factor receptor in lung cancer

2015 ◽  
Vol 26 (22) ◽  
pp. 4087-4099 ◽  
Author(s):  
Christopher C. Valley ◽  
Donna J. Arndt-Jovin ◽  
Narain Karedla ◽  
Mara P. Steinkamp ◽  
Alexey I. Chizhik ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Epidermal Growth Factor Receptor ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Molecular Mechanisms ◽  
Critical Role ◽  
Resonance Energy Transfer ◽  
Growth Factor Receptor ◽  
Resonance Energy ◽  
Epidermal Growth

Mutations within the epidermal growth factor receptor (EGFR/erbB1/Her1) are often associated with tumorigenesis. In particular, a number of EGFR mutants that demonstrate ligand-independent signaling are common in non–small cell lung cancer (NSCLC), including kinase domain mutations L858R (also called L834R) and exon 19 deletions (e.g., ΔL747-P753insS), which collectively make up nearly 90% of mutations in NSCLC. The molecular mechanisms by which these mutations confer constitutive activity remain unresolved. Using multiple subdiffraction-limit imaging modalities, we reveal the altered receptor structure and interaction kinetics of NSCLC-associated EGFR mutants. We applied two-color single quantum dot tracking to quantify receptor dimerization kinetics on living cells and show that, in contrast to wild-type EGFR, mutants are capable of forming stable, ligand-independent dimers. Two-color superresolution localization microscopy confirmed ligand-independent aggregation of EGFR mutants. Live-cell Förster resonance energy transfer measurements revealed that the L858R kinase mutation alters ectodomain structure such that unliganded mutant EGFR adopts an extended, dimerization-competent conformation. Finally, mutation of the putative dimerization arm confirmed a critical role for ectodomain engagement in ligand-independent signaling. These data support a model in which dysregulated activity of NSCLC-associated kinase mutants is driven by coordinated interactions involving both the kinase and extracellular domains that lead to enhanced dimerization.

Monitoring the Structural Dynamics of U2 snRNA Via Single-Molecule Förster Resonance Energy Transfer

2018 ◽  
Author(s):  
Alexander Carl DeHaven
Keyword(s):  
Energy Transfer ◽  
Structural Dynamics ◽  
Single Molecule ◽  
Conformational Dynamics ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Förster Resonance Energy Transfer ◽  
U2 Snrna ◽  
Folding Dynamics ◽  
The Impact

This thesis contains four topic areas: a review of single-molecule microscropy methods and splicing, conformational dynamics of stem II of the U2 snRNA, the impact of post-transcriptional modifications on U2 snRNA folding dynamics, and preliminary findings on Mango aptamer folding dynamics.

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