ArticleAnalytical biochemistry2018
In situ measurements of mitochondrial matrix enzyme activities using plasma and mitochondrial membrane permeabilization agents.
Article in Analytical biochemistry, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 16 citations in OpenAlex.
- Small-molecule binding-site discovery using silyl ether-enabled chemoproteomics.Nature chemistry · 2026Article
- Pro-inflammatory macrophage activation does not require inhibition of oxidative phosphorylation.EMBO reports · 2025Article
- Exogenous expression of ATP8, a mitochondrial encoded protein, from the nucleusMolecular therapy. Methods & clinical development · 2024Article
- Pro-inflammatory macrophage activation does not require inhibition of mitochondrial respiration.bioRxiv : the preprint server for biology · 2024Article
- A novel approach to measure complex V ATP hydrolysis in frozen cell lysates and tissue homogenates.Life science alliance · 2023Article
- A practical guide for the analysis, standardization and interpretation of oxygen consumption measurements.Nature metabolism · 2022Review
- Utilization of Human Samples for Assessment of Mitochondrial Bioenergetics: Gold Standards, Limitations, and Future Perspectives.Life (Basel, Switzerland) · 2021Review
- Mapping mitochondrial respiratory chain deficiencies by respirometry: Beyond the Mito Stress Test.Experimental neurology · 2020Article
- Etomoxir Inhibits Macrophage Polarization by Disrupting CoA Homeostasis.Cell metabolism · 2018Article
Corrections and comments
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Authors and funding
4 authors at 3 institutions in 1 country.
Funding
Abstract
Activities of enzymes localized to the mitochondrial matrix of mammalian cells are often critical regulatory steps in cellular metabolism. As such, measurement of matrix enzyme activities in response to genetic modifications or drug interventions is often desired. However, measurements in intact cells are often hampered by the presence of other isozymes in the cytoplasm as well as the inability to deliver enzyme substrates across cellular membranes. Classic approaches to liberate matrix enzymes utilize harsh treatments that disrupt intracellular architecture or require significant starting material to allow mitochondrial isolation prior to sample extraction. We describe a method using permeabilization reagents for both the plasma and mitochondrial membranes to allow in situ measurement of matrix enzyme activities. It is applied to adherent cell monolayers in 96-well plates treated with perfringolysin O to permeabilize the plasma membrane and alamethicin to permeabilize the mitochondrial inner membrane. We present three examples validated with inhibitor sensitivity: (i) Complex I-mediated oxygen consumption driven by NADH, (ii) ATP hydrolysis by the F
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.