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

Mitochondrial Membranes01:45

Mitochondrial Membranes

17.7K
A single mitochondrion is a bean-shaped organelle enclosed by a double-membrane system. The outer membrane of mitochondria is smooth and contains many porins - the integral membrane transporters. Porins enable free diffusion of ions and small uncharged molecules through the outer mitochondrial membrane but limit the transport of molecules larger than 5000 Daltons. Further, the outer mitochondrial membrane forms a unique structure called membrane contact sites with other subcellular organelles,...
17.7K
Translocation of Proteins into the Mitochondria01:19

Translocation of Proteins into the Mitochondria

13.6K
Mitochondrial precursors are translocated to the internal subcompartments via independent mechanisms involving distinct protein machineries called translocases.
Sorting of outer membrane proteins:
Mitochondrial outer membrane proteins are of two types: the transmembrane, beta-barrel porins, and the membrane-anchored, alpha-helical proteins. Beta-barrel porin precursors are translocated by the TOM complex and inserted into the outer mitochondrial membrane by the SAM complex. In contrast,...
13.6K

You might also read

Related Articles

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

Sort by
Same author

Biophysical and structural analysis of human green cone opsin.

Biophysical journal·2026
Same author

Structure of human green cone opsin yields insights into mechanisms underlying the rapid decay of its active, signaling state.

Proceedings of the National Academy of Sciences of the United States of America·2025
Same author

A novel "bio-tag" for cryo-EM studies based on the small, electron-dense protein Csp1.

Biophysical journal·2025
Same author

Active and inactive pathways in the kinetic mechanism of the G51V retinitis pigmentosa mutant photoreaction.

Biophysical journal·2025
Same author

A rapid, tag-free way to purify functional GPCRs.

The Journal of biological chemistry·2023
Same author

Styrene-maleic acid copolymer effects on the function of the GPCR rhodopsin in lipid nanoparticles.

Biophysical journal·2021

Related Experiment Video

Updated: Mar 19, 2026

Author Spotlight: Advancing Mitochondrial Research - mtHyper7 Biosensor for Subcellular Analysis
09:47

Author Spotlight: Advancing Mitochondrial Research - mtHyper7 Biosensor for Subcellular Analysis

Published on: June 2, 2023

3.5K

Biosensor reveals multiple sources for mitochondrial NAD⁺.

Xiaolu A Cambronne1, Melissa L Stewart1, DongHo Kim1

  • 1Vollum Institute, Oregon Health & Science University, Portland, OR 97239, USA.

Science (New York, N.Y.)
|June 18, 2016
PubMed
Summary

Researchers developed a novel biosensor to measure cellular nicotinamide adenine dinucleotide (NAD(+)) levels. This tool revealed compartmentalized NAD(+) concentrations regulate key enzymes, with cells employing unique strategies to maintain mitochondrial NAD(+).

More Related Videos

Simultaneous Measurement of Superoxide/Hydrogen Peroxide and NADH Production by Flavin-containing Mitochondrial Dehydrogenases
08:57

Simultaneous Measurement of Superoxide/Hydrogen Peroxide and NADH Production by Flavin-containing Mitochondrial Dehydrogenases

Published on: February 24, 2018

10.6K
Ratiometric Biosensors that Measure Mitochondrial Redox State and ATP in Living Yeast Cells
12:22

Ratiometric Biosensors that Measure Mitochondrial Redox State and ATP in Living Yeast Cells

Published on: July 22, 2013

21.8K

Related Experiment Videos

Last Updated: Mar 19, 2026

Author Spotlight: Advancing Mitochondrial Research - mtHyper7 Biosensor for Subcellular Analysis
09:47

Author Spotlight: Advancing Mitochondrial Research - mtHyper7 Biosensor for Subcellular Analysis

Published on: June 2, 2023

3.5K
Simultaneous Measurement of Superoxide/Hydrogen Peroxide and NADH Production by Flavin-containing Mitochondrial Dehydrogenases
08:57

Simultaneous Measurement of Superoxide/Hydrogen Peroxide and NADH Production by Flavin-containing Mitochondrial Dehydrogenases

Published on: February 24, 2018

10.6K
Ratiometric Biosensors that Measure Mitochondrial Redox State and ATP in Living Yeast Cells
12:22

Ratiometric Biosensors that Measure Mitochondrial Redox State and ATP in Living Yeast Cells

Published on: July 22, 2013

21.8K

Area of Science:

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Background:

  • Nicotinamide adenine dinucleotide (NAD(+)) is crucial for cellular metabolism and DNA repair.
  • NAD(+)-consuming enzymes like sirtuins and PARPs are vital but their activity regulation by compartmentalized NAD(+) remains unclear.
  • Measuring subcellular NAD(+) levels has been a significant technical challenge.

Purpose of the Study:

  • To develop a genetically encoded fluorescent biosensor for real-time monitoring of free NAD(+) concentrations in distinct subcellular compartments.
  • To investigate the relationship between compartmentalized NAD(+) levels and the activity of NAD(+)-consuming enzymes.
  • To elucidate cellular mechanisms governing mitochondrial NAD(+) homeostasis.

Main Methods:

  • Development of a genetically encoded fluorescent biosensor for intracellular NAD(+) detection.
  • Utilizing the biosensor to measure free NAD(+) concentrations in the nucleus, cytoplasm, and mitochondria.
  • Employing genetic depletion of NAD(+) biosynthesis enzymes to study mitochondrial NAD(+) maintenance.

Main Results:

  • The biosensor successfully enabled direct measurement of free NAD(+) in subcellular compartments.
  • Measured nuclear, cytoplasmic, and mitochondrial NAD(+) concentrations were found to be near the Michaelis constants of key NAD(+)-consuming enzymes.
  • Depletion of NAD(+) biosynthesis enzymes highlighted cell-specific strategies for maintaining mitochondrial NAD(+) levels.

Conclusions:

  • Compartmentalized NAD(+) concentrations directly influence the activity of sirtuins and PARPs.
  • The developed biosensor provides a powerful tool for studying NAD(+) metabolism and signaling in living cells.
  • Cellular NAD(+) homeostasis, particularly in mitochondria, is maintained through sophisticated, cell-type-specific regulatory mechanisms.