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 Concept Videos

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

2.1K
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
2.1K

You might also read

Related Articles

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

Sort by
Same author

Comparative Evaluation of the Clinical Performance of Two-step and One-step Self-etching Adhesive Systems and Self-etching Self-adhesive Composite in Permanent Molars in Pediatric Patients: A Three-arm Parallel Randomized Clinical Study.

International journal of clinical pediatric dentistry·2026
Same author

Di-ortho-fluoroazobenzene Functionalized Discotic Nematic Liquid Crystals as Soft Ambipolar Semiconductors.

Chemistry, an Asian journal·2026
Same author

Pd-Decorated Ag-CeO<sub>2</sub>/PANI Nanocomposite: Fabrication, Application in the Synthesis of 2,3-Dihydroquinazolin-4(1<i>H</i>)-ones and Tetrahydrobenzo[<i>b</i>]pyrans, Antimicrobial Activity Evaluation, and Computational Study.

ACS applied bio materials·2026
Same author

Comparison of human vs AI-generated feedback: results from an online survey study in the GlomCon glomerular disease fellowship program.

Journal of nephrology·2026
Same author

Evolution of incretin-based therapies: From GLP-1 monotherapy to dual and triple agonists: A new era in metabolic therapy.

The Indian journal of medical research·2026
Same author

Hydraulic geometry hypothesis allows reverse engineering of 3D quasi-equilibrium landscapes from 2D channel networks: Earth, Mars, Titan.

Proceedings of the National Academy of Sciences of the United States of America·2026

Related Experiment Video

Updated: Jun 25, 2025

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules
00:10

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules

Published on: September 5, 2019

8.2K

Deeper Insights into Mixed Crowding through Enzyme Activity, Dynamics, and Crowder Diffusion.

Arvind Singh1, Monika Gupta1, Harshita Rastogi1

  • 1Department of Chemistry, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India.

The Journal of Physical Chemistry. B
|May 29, 2024
PubMed
Summary

Cellular crowding affects enzyme function. Binary crowder mixtures reduce adenylate kinase activity and dynamics, with enzymes avoiding dense, crowder-rich droplets, unlike in single crowder systems.

More Related Videos

Single-Molecule Diffusion and Assembly on Polymer-Crowded Lipid Membranes
10:43

Single-Molecule Diffusion and Assembly on Polymer-Crowded Lipid Membranes

Published on: July 19, 2022

2.3K
Characterizing Microbiome Dynamics &#8211; Flow Cytometry Based Workflows from Pure Cultures to Natural Communities
09:57

Characterizing Microbiome Dynamics – Flow Cytometry Based Workflows from Pure Cultures to Natural Communities

Published on: July 12, 2018

11.9K

Related Experiment Videos

Last Updated: Jun 25, 2025

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules
00:10

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules

Published on: September 5, 2019

8.2K
Single-Molecule Diffusion and Assembly on Polymer-Crowded Lipid Membranes
10:43

Single-Molecule Diffusion and Assembly on Polymer-Crowded Lipid Membranes

Published on: July 19, 2022

2.3K
Characterizing Microbiome Dynamics &#8211; Flow Cytometry Based Workflows from Pure Cultures to Natural Communities
09:57

Characterizing Microbiome Dynamics – Flow Cytometry Based Workflows from Pure Cultures to Natural Communities

Published on: July 12, 2018

11.9K

Area of Science:

  • Biochemistry
  • Biophysics
  • Cellular Biology

Background:

  • Cellular interiors are crowded with macromolecules, influencing biomolecular behavior.
  • In vitro studies require mimicking this crowded environment using crowder systems.

Purpose of the Study:

  • To investigate the effects of binary crowder mixtures on enzyme activity, dynamics, and diffusion.
  • To understand the simplest form of mixed crowding and its implications for cellular complexity.

Main Methods:

  • Used binary mixtures of Ficoll 70, Dextran 70/40, and PEG 8000 as crowders.
  • Studied adenylate kinase 1-like 1 (AK3L1) enzyme activity and dynamics.
  • Employed single point mutations and coumarin-based solvatochromic probe (CPM) labeling for dynamics.
  • Utilized fluorescence correlation spectroscopy (FCS) with labeled crowders and enzyme.
  • Performed confocal and digital holographic microscopy for imaging phase-separated droplets.

Main Results:

  • Binary crowder mixtures reduced enzyme activity and dynamics compared to individual crowders.
  • Enzyme diffusion was more constrained than crowder diffusion in binary mixtures.
  • Phase-separated droplets formed in PEG-containing mixtures were crowder-rich.
  • Enzymes preferentially localized outside these dense, crowder-rich droplets.

Conclusions:

  • Binary crowder systems alter enzyme behavior, reducing activity and dynamics.
  • Enzymes and crowders experience different environments within binary mixtures.
  • Phase separation in mixed crowders leads to spatial segregation of enzymes.
  • These findings highlight the complexity of macromolecular crowding in cellular environments.