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

Rab Cascades01:25

Rab Cascades

3.8K
Rab GTPases act in a regulated cascade during membrane fusion, helping the lipid bilayers mix. The Rab family of proteins are active when bound to GTP, and inactive when bound to GDP. Hence, they act as guanine nucleotide-dependent molecular switches. Rab-GTP recognizes and binds to long or short-range tethering proteins to capture the target vesicle. These tethers coordinate with SNAREs on the vesicle and the target membrane to assemble the trans SNARE complex that locks the mixing bilayers.
3.8K
Rab Proteins01:14

Rab Proteins

5.5K
Rab proteins constitute the largest family of monomeric GTPases, of which 70 members are present in humans. Rab proteins and their effectors regulate consecutive stages of vesicle transport such as vesicle transport, docking, and fusion to the correct recipient membrane.
Rab proteins switch between a cytosolic, GDP-bound inactive state and a membrane-anchored, GTP-bound active state. By themselves, Rabs show slow rates of GDP/GTP exchange and GTP hydrolysis. Thus, Rab proteins are considered...
5.5K
Bacterial Translocation and Protein Secretion01:26

Bacterial Translocation and Protein Secretion

1.0K
Bacterial protein secretion involves translocation systems to ensure proteins reach their designated locations, including the plasma membrane, periplasm, outer membrane, or the external environment. These translocation systems are vital for bacterial physiology, supporting processes like membrane assembly, enzymatic activity in the periplasm, and interactions with the external environment. The division of labor between Sec and Tat pathways ensures efficiency in handling proteins with diverse...
1.0K
Intralumenal Vesicles and Multivesicular Bodies01:38

Intralumenal Vesicles and Multivesicular Bodies

5.3K
Intraluminal vesicles (ILVs) are small vesicles 50-80 nm in diameter formed during the maturation of early endosomes. A specialized endosome containing numerous ILVs is called a multivesicular body (MVB). ILVs contain internalized molecules such as antigens, nucleic acids, proteins, and metabolites. Some of these molecules are released from the MVBs inside exosomes and are transported to other cells. Other MVBs contain molecules that are retained in the ILVs and are later degraded within the...
5.3K
Cotranslational Protein Translocation01:20

Cotranslational Protein Translocation

11.1K
Translocation of proteins across membranes is an ancient process that occurs even in bacteria and archaebacteria. In fact, the components of the translocation machinery are still conserved between prokaryotes and eukaryotes.
Sec61 channel partners for cotranslational translocation
During cotranslational translocation, the Sec61 channel partners with the signal recognition particle (SRP), the signal recognition particle receptor (SR), and the ribosomes to transport the nascent polypeptide chain...
11.1K
Post-translational Translocation of Proteins to the RER01:27

Post-translational Translocation of Proteins to the RER

8.2K
A sizable fraction of proteins destined for ER are first synthesized in the cell cytosol and then transported across the ER membrane–a process called post-translational translocation. Similar to cotranslationally translocated proteins, these proteins also use the Sec translocon complex to enter the ER lumen.
Targeting proteins to the ER
Hsp40 and Hsp70 chaperone molecules bind the translated proteins in the cytosol to prevent their folding. The chaperone binding helps to keep the signal...
8.2K

You might also read

Related Articles

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

Sort by
Same author

Relative importance of the anti-apoptotic versus apoptosis-unrelated functions of MCL-1 in vivo.

Science (New York, N.Y.)·2025
Same author

The diagnostic pathway of Parkinson's disease: understanding patient perspectives in Australia.

NPJ Parkinson's disease·2025
Same author

Differential regulation of BAX and BAK apoptotic activity revealed by small molecules.

Science advances·2025
Same author

Mitochondria and cell death.

Nature cell biology·2024
Same author

Key residues in the VDAC2-BAK complex can be targeted to modulate apoptosis.

PLoS biology·2024
Same author

Interaction of PINK1 with nucleotides and kinetin.

Science advances·2024
Same journal

Emerging Trends in Mass Spectrometry-Based Quantitative Proteome and Phosphoproteome Profiling in Maize.

Cold Spring Harbor protocols·2026
Same journal

Sample Preparation for Quantitative Proteome and Phosphoproteome Profiling of Maize Tissues.

Cold Spring Harbor protocols·2026
Same journal

High-Throughput Microbial Assay for Amino Acid Measurement in Ground Maize Seed Samples Utilizing Auxotrophic <i>E. coli</i>.

Cold Spring Harbor protocols·2025
Same journal

Grain Quality in Maize.

Cold Spring Harbor protocols·2025
Same journal

High-Throughput Assay for Measuring Phytate and Available Phosphorus in Ground Maize Seed Samples.

Cold Spring Harbor protocols·2025
Same journal

Functional Genomic Analysis of Transposon Insertion Mutant Maize Plants from the UniformMu National Public Resource.

Cold Spring Harbor protocols·2025
See all related articles

Related Experiment Video

Updated: Apr 13, 2026

Examining BCL-2 Family Function with Large Unilamellar Vesicles
08:35

Examining BCL-2 Family Function with Large Unilamellar Vesicles

Published on: October 5, 2012

8.5K

Investigating Bax subcellular localization and membrane integration.

Grant Dewson1

  • 1Cell Signalling and Cell Death Division, Walter and Eliza Hall Institute of Medical Research, Parkville, Melbourne, Victoria 3052, Australia;Department of Medical Biology, The University of Melbourne, Melbourne, Victoria 3010, Australia.

Cold Spring Harbor Protocols
|May 3, 2015
PubMed
Summary
This summary is machine-generated.

This study introduces digitonin fractionation, a simple method to track Bax protein localization during apoptosis. It differentiates Bax in the cytosol versus mitochondria and distinguishes integrated from peripheral membrane-associated Bax.

More Related Videos

Purification of the Membrane Compartment for Endoplasmic Reticulum-associated Degradation of Exogenous Antigens in Cross-presentation
12:48

Purification of the Membrane Compartment for Endoplasmic Reticulum-associated Degradation of Exogenous Antigens in Cross-presentation

Published on: August 21, 2017

8.7K
Expression, Purification, and Liposome Binding of Budding Yeast SNX-BAR Heterodimers
10:28

Expression, Purification, and Liposome Binding of Budding Yeast SNX-BAR Heterodimers

Published on: December 6, 2019

8.3K

Related Experiment Videos

Last Updated: Apr 13, 2026

Examining BCL-2 Family Function with Large Unilamellar Vesicles
08:35

Examining BCL-2 Family Function with Large Unilamellar Vesicles

Published on: October 5, 2012

8.5K
Purification of the Membrane Compartment for Endoplasmic Reticulum-associated Degradation of Exogenous Antigens in Cross-presentation
12:48

Purification of the Membrane Compartment for Endoplasmic Reticulum-associated Degradation of Exogenous Antigens in Cross-presentation

Published on: August 21, 2017

8.7K
Expression, Purification, and Liposome Binding of Budding Yeast SNX-BAR Heterodimers
10:28

Expression, Purification, and Liposome Binding of Budding Yeast SNX-BAR Heterodimers

Published on: December 6, 2019

8.3K

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Bax protein is crucial for initiating apoptosis by permeabilizing the mitochondrial outer membrane (MOM).
  • Understanding Bax translocation to the MOM is key to studying mitochondrial damage during apoptosis.

Purpose of the Study:

  • To present a straightforward and effective method for assessing Bax localization between the cytosol and mitochondria.
  • To differentiate between membrane-integrated and peripherally associated Bax using carbonate extraction.

Main Methods:

  • Digitonin fractionation to separate cytosolic and heavy membrane fractions (containing mitochondria and nuclei).
  • Differential ultracentrifugation to isolate cellular compartments.
  • Immunoblotting to detect Bax protein.
  • High pH carbonate extraction to distinguish integrated from peripheral membrane proteins.

Main Results:

  • Digitonin fractionation effectively separates Bax in the cytosol from Bax associated with the MOM.
  • Carbonate extraction differentiates integrated Bax from peripherally associated Bax in the membrane fraction.

Conclusions:

  • Digitonin fractionation is a valuable technique for studying Bax localization dynamics.
  • This method aids in understanding the molecular mechanisms of Bax-mediated apoptosis and mitochondrial dysfunction.