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 Folding01:22

Protein Folding

130.7K
Overview
130.7K
Protein Folding01:25

Protein Folding

12.6K
Proteins are chains of amino acids linked together by peptide bonds. Upon synthesis, a protein folds into a three-dimensional conformation, critical to its biological function. Interactions between its constituent amino acids guide protein folding, and hence the protein structure is primarily dependent on its amino acid sequence.
Protein Structure Is Critical to Its Biological Function
Proteins perform a wide range of biological functions such as catalyzing chemical reactions, providing...
12.6K
Bacterial Protein Maturation01:26

Bacterial Protein Maturation

705
Bacterial protein maturation is a tightly regulated process that ensures newly synthesized polypeptides achieve correct functional conformations. This maturation involves a series of modifications, folding events, and quality control steps, often assisted by specialized chaperone proteins.N-Terminal ModificationsThe maturation of bacterial polypeptides begins cotranslationally as the polypeptide exits the ribosome. The first amino acid, N-formylmethionine (fMet), is typically modified at the...
705
Protein Modifications in the RER01:26

Protein Modifications in the RER

7.6K
Modification of secretory and transmembrane proteins entering the rough ER begins in the ER lumen. These modifications aid in protein folding and stabilize the acquired tertiary structure. Protein modifications in the rough ER co-occur at different stages of protein folding.
Broadly, these modifications can be categorized into four main categories — glycosylation, formation of disulfide bonds, assembly of protein subunits, and specific proteolytic cleavages like removal of signal...
7.6K
Covalently Linked Protein Regulators02:04

Covalently Linked Protein Regulators

9.9K
Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein....
9.9K

You might also read

Related Articles

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

Sort by
Same author

Admissions to MD-PhD programs: how well do application metrics predict short- or long-term physician-scientist outcomes?

JCI insight·2025
Same author

The financial impact of MD-PhD training compared with MD training for academic physicians.

JCI insight·2024
Same author

Yeast-based assay to identify inhibitors of the malaria parasite sodium phosphate uptake transporter as potential novel antimalarial drugs.

International journal for parasitology. Drugs and drug resistance·2024
Same author

More women than ever are entering MD-PhD programs. What lies ahead for them?

JCI insight·2024
Same author

The National MD-PhD Program Outcomes Study: career paths followed by Black and Hispanic graduates.

JCI insight·2024
Same author

Sociodemographic factors and research experience impact MD-PhD program acceptance.

JCI insight·2024

Related Experiment Video

Updated: Apr 3, 2026

Examining the Conformational Dynamics of Membrane Proteins in situ with Site-directed Fluorescence Labeling
11:55

Examining the Conformational Dynamics of Membrane Proteins in situ with Site-directed Fluorescence Labeling

Published on: May 29, 2011

15.6K

Cysteine Modification: Probing Channel Structure, Function and Conformational Change.

Myles H Akabas1

  • 1Departments of Physiology & Biophysics, Neuroscience and Medicine, Albert Einstein College of Medicine, 1300 Morris Park Avenue, 10461, Bronx, NY, USA. myles.akabas@einstein.yu.edu.

Advances in Experimental Medicine and Biology
|September 19, 2015
PubMed
Summary
This summary is machine-generated.

Cysteine mutagenesis is a key method for studying membrane protein structure and function. Engineered cysteines allow detailed probing of protein environments and conformational changes, especially in ion channels and Cys-loop receptors.

Keywords:
Acetylcholine receptorGABAA receptorMembrane transporterMethanethiosulfonatePotassium channelsSCAMSulfhydrylThiolThiolate

More Related Videos

Targeting Cysteine Thiols for in Vitro Site-specific Glycosylation of Recombinant Proteins
11:25

Targeting Cysteine Thiols for in Vitro Site-specific Glycosylation of Recombinant Proteins

Published on: October 4, 2017

7.2K
Author Spotlight: Functional Site-Directed Fluorometry in Native Cells to Study Skeletal Muscle Excitability
12:26

Author Spotlight: Functional Site-Directed Fluorometry in Native Cells to Study Skeletal Muscle Excitability

Published on: June 2, 2023

1.6K

Related Experiment Videos

Last Updated: Apr 3, 2026

Examining the Conformational Dynamics of Membrane Proteins in situ with Site-directed Fluorescence Labeling
11:55

Examining the Conformational Dynamics of Membrane Proteins in situ with Site-directed Fluorescence Labeling

Published on: May 29, 2011

15.6K
Targeting Cysteine Thiols for in Vitro Site-specific Glycosylation of Recombinant Proteins
11:25

Targeting Cysteine Thiols for in Vitro Site-specific Glycosylation of Recombinant Proteins

Published on: October 4, 2017

7.2K
Author Spotlight: Functional Site-Directed Fluorometry in Native Cells to Study Skeletal Muscle Excitability
12:26

Author Spotlight: Functional Site-Directed Fluorometry in Native Cells to Study Skeletal Muscle Excitability

Published on: June 2, 2023

1.6K

Area of Science:

  • Biochemistry and Molecular Biology
  • Structural Biology
  • Membrane Protein Research

Background:

  • Cysteine's unique reactivity makes it valuable for protein studies.
  • Engineered cysteines allow site-specific labeling and probing within proteins.
  • Membrane proteins are crucial but challenging to study in their native environment.

Purpose of the Study:

  • To review the application of cysteine mutagenesis in understanding membrane protein structure-function relationships.
  • To highlight the utility of cysteine substitution for probing ion channels and Cys-loop receptors.
  • To emphasize how cysteine accessibility and disulfide bond formation reveal protein dynamics.

Main Methods:

  • Site-directed mutagenesis to introduce cysteine residues at specific protein locations.
  • Utilizing sulfhydryl-reactive reagents for labeling engineered cysteines.
  • Employing the Substituted Cysteine Accessibility Method (SCAM) for probing protein environments.
  • Analyzing disulfide bond formation between introduced cysteine pairs to infer proximity and mobility.

Main Results:

  • Cysteine substitution is generally well-tolerated, enabling systematic studies of various protein regions.
  • SCAM successfully identifies residues within ion channels, binding sites, and conformational change regions.
  • Cysteine accessibility and disulfide mapping provide insights into protein structure and dynamics in situ.
  • Data complements structural information from crystal structures of detergent-solubilized proteins.

Conclusions:

  • Cysteine mutagenesis is a powerful and versatile tool for investigating membrane protein structure and function.
  • It provides critical insights into the functional environment and dynamic behavior of proteins like ion channels and receptors.
  • This technique offers complementary data to traditional structural biology methods, particularly for membrane proteins.