Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Video

Updated: Jun 3, 2026

Generation of Alpha-Synuclein Preformed Fibrils from Monomers and Use In Vivo
09:44

Generation of Alpha-Synuclein Preformed Fibrils from Monomers and Use In Vivo

Published on: June 2, 2019

Site-Specific Raman Probes Reveal Droplet Aging and Residue-Level Fibril Polymorphism in TDP-43CTD.

Matthew D Watson1, Jennifer C Lee1

  • 1Laboratory of Protein Conformation and Dynamics, Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, United States.

Journal of the American Chemical Society
|June 2, 2026
PubMed
Summary

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Kinetics and Spatial Distribution of β-Sheet Development in TDP-43<sub>CTD</sub> Condensate Maturation.

ACS chemical neuroscience·2026
Same author

A dual fluorescent-Raman bioorthogonal probe for specific biosynthetic labeling of intracellular gangliosides.

Communications chemistry·2025
Same author

Amyloid formation of alternatively spliced variants of α-synuclein.

Protein science : a publication of the Protein Society·2025
Same author

A Dual Fluorescent-Raman Bioorthogonal Probe for Specific Biosynthetic Labeling of Gangliosides.

Research square·2025
Same author

Defining essential charged residues in fibril formation of a lysosomal derived N-terminal α-synuclein truncation.

Nature communications·2025
Same author

Raman spectroscopy in the study of amyloid formation and phase separation.

Biochemical Society transactions·2024

Researchers developed a new method to study how proteins like TDP-43CTD change from liquid droplets to amyloid aggregates, a key process in neurodegenerative diseases. This technique reveals distinct structural changes at the single-residue level.

Area of Science:

  • Biochemistry
  • Biophysics
  • Neuroscience

Background:

  • The C-terminal domain of TAR DNA-binding protein 43 (TDP-43CTD) is central to liquid-liquid phase separation (LLPS) and amyloid formation.
  • The transition of TDP-43CTD droplets into amyloid aggregates is implicated in neurodegenerative diseases like ALS and FTD.
  • Site-specific structural characterization of this transition at high spatial resolution is crucial for understanding disease mechanisms.

Purpose of the Study:

  • To develop and apply a novel methodology for site-specific structural characterization of TDP-43CTD droplets during aging and amyloid formation.
  • To investigate the conformational changes occurring within individual TDP-43CTD droplets using confocal Raman spectroscopy and alkyne-modified amino acids.
  • To elucidate the relationship between LLPS, droplet aging, and amyloid aggregation at the single-residue level.

More Related Videos

Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy
14:04

Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy

Published on: April 25, 2021

Characterizing Individual Protein Aggregates by Infrared Nanospectroscopy and Atomic Force Microscopy
12:58

Characterizing Individual Protein Aggregates by Infrared Nanospectroscopy and Atomic Force Microscopy

Published on: September 12, 2019

Related Experiment Videos

Last Updated: Jun 3, 2026

Generation of Alpha-Synuclein Preformed Fibrils from Monomers and Use In Vivo
09:44

Generation of Alpha-Synuclein Preformed Fibrils from Monomers and Use In Vivo

Published on: June 2, 2019

Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy
14:04

Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy

Published on: April 25, 2021

Characterizing Individual Protein Aggregates by Infrared Nanospectroscopy and Atomic Force Microscopy
12:58

Characterizing Individual Protein Aggregates by Infrared Nanospectroscopy and Atomic Force Microscopy

Published on: September 12, 2019

Main Methods:

  • Incorporation of an alkyne-modified amino acid (4-ethynyl-l-phenylalanine, FCC) into TDP-43CTD.
  • Site-specific probing of protein structure within individual TDP-43CTD droplets using confocal Raman spectroscopy.
  • Analysis of spectral changes in the alkyne stretching band to differentiate between disordered, β-sheet, and amyloid states.

Main Results:

  • Nascent TDP-43CTD droplets are initially disordered, with β-sheet conformers developing in aged droplets and amyloid aggregates.
  • The alkyne stretching band provides distinct spectral signatures for disordered, β-sheet, and amyloid states, with site-dependent sensitivity.
  • C-terminal sites (Y374FCC, W385FCC, F397FCC) act as sensitive probes for amyloid polymorphs, revealing heterogeneity not seen with global methods.
  • W334FCC incorporation inhibited β-sheet formation, yet *de novo* aggregation still occurred, decoupling droplet aging from amyloid formation.

Conclusions:

  • This study establishes a generalizable approach for investigating protein conformational changes during LLPS and amyloid formation *in cellulo*.
  • The methodology allows for high-resolution, site-specific characterization of protein aggregation pathways.
  • Droplet aging is not a prerequisite for amyloid formation in TDP-43CTD, challenging previous assumptions.