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

You might also read

Related Articles

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

Sort by
Same author

Phospholipid-driven conformational switching of HCV NS5A links protein folding to replication membrane remodeling.

Science advances·2026
Same author

Single-molecule dynamics reveal ATP binding alone powers substrate translocation by an ABC transporter.

Nature communications·2026
Same author

Braess' Paradox in Enzyme Kinetics: Asymmetry from Population Balance without Direct Cooperativity.

Journal of chemical theory and computation·2026
Same author

Architectural principles of transporter-chaperone coupling within the native MHC I peptide-loading complex.

Science advances·2026
Same author

Dendritic cell phagosomes recruit GRASP55 for export of antigen-loaded MHC molecules.

Cell reports·2025
Same author

Engineering Mesoscale T Cell Receptor Clustering by Plug-and-Play Nanotools.

Advanced materials (Deerfield Beach, Fla.)·2024
Same journal

Clinical Europium fluorescent based lectin assays for mucin O-glycomics.

Methods in enzymology·2026
Same journal

A dual-color FRET assay for detection and quantitative analysis of O-glycopeptidases.

Methods in enzymology·2026
Same journal

Evolutionary genetic approaches to analyze mucins.

Methods in enzymology·2026
Same journal

Ex vivo imaging and enzymatic analysis of intestinal mucus.

Methods in enzymology·2026
Same journal

Glyco-TRAPP: A real-time glycocalyx permeability assay for assessing transmembrane mucin barrier function in live and fixed tissues.

Methods in enzymology·2026
Same journal

Quantitative imaging approaches to capture structural and functional dynamics of colonic mucus in health and disease in situ.

Methods in enzymology·2026
See all related articles

Related Experiment Video

Updated: Apr 12, 2026

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli
08:46

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

Published on: January 6, 2015

33.8K

Multicolor fluorescence-based screening toward structural analysis of multiprotein membrane complexes.

Simon Trowitzsch1, Robert Tampé2

  • 1Institute of Biochemistry, Biocenter, Goethe-University Frankfurt, Frankfurt/Main, Germany.

Methods in Enzymology
|May 8, 2015
PubMed
Summary
This summary is machine-generated.

Determining membrane protein complex structures is challenging due to low native abundance. This study introduces a fluorescence-based screening method to optimize production and purification for structural studies.

Keywords:
ABC transportersFluorescence-based screeningHeterologous expression systemsMulticolor fluorescence-detection size-exclusion chromatographyMultisubunit membrane protein complexesPeptide-loading complexStructural biologyTransporter associated with antigen processing

More Related Videos

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
11:06

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

9.2K
Rapid Assessment of Membrane Protein Quality by Fluorescent Size Exclusion Chromatography
06:26

Rapid Assessment of Membrane Protein Quality by Fluorescent Size Exclusion Chromatography

Published on: January 6, 2023

4.2K

Related Experiment Videos

Last Updated: Apr 12, 2026

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli
08:46

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

Published on: January 6, 2015

33.8K
Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
11:06

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

9.2K
Rapid Assessment of Membrane Protein Quality by Fluorescent Size Exclusion Chromatography
06:26

Rapid Assessment of Membrane Protein Quality by Fluorescent Size Exclusion Chromatography

Published on: January 6, 2023

4.2K

Area of Science:

  • Structural biology
  • Biochemistry
  • Membrane protein research

Background:

  • Membrane protein complexes are crucial for biological functions, but their structural determination is hindered by low native abundance.
  • Heterologous expression systems are vital for overproducing these complexes, yet challenges like poor expression and instability persist.
  • Efficient tools are needed to guide the selection of expression hosts and optimize purification strategies for membrane protein complexes.

Purpose of the Study:

  • To present a novel fluorescence-based screening approach for analyzing the production and purification of multiprotein membrane complexes.
  • To enable sensitive and rapid assessment of critical subunits during the production process.
  • To facilitate optimization for monodispersity, stoichiometry, and stability of membrane protein complexes.

Main Methods:

  • Development and application of a multicolor fluorescence-detection size-exclusion chromatography (SEC) system.
  • Quantitative monitoring of individual subunit production during heterologous expression.
  • Tracking sample quality and stability throughout purification.

Main Results:

  • The fluorescence-based SEC approach allows sensitive detection and quantification of membrane protein complex subunits.
  • The method effectively monitors production yields and identifies critical bottlenecks in expression and purification.
  • Optimization strategies can be rapidly assessed, leading to improved complex monodispersity, stoichiometry, and stability.

Conclusions:

  • Fluorescence-detection SEC is a powerful tool for optimizing the production and purification of challenging membrane protein complexes.
  • This screening approach accelerates structural genomics efforts by enabling efficient sample preparation.
  • The methodology supports the structural characterization of membrane protein complexes previously inaccessible due to production difficulties.