Mitochondria and mitochondrial research

Research tools aimed at exploring mitochondria are becoming more and more popular and cover multiple experimental applications. They go from primary antibodies validated for immuno-assays (see “10 Mitochondrial marker antibodies“) to fluorescent probes for mitochondria-related events monitor in living-cells (e.g. MitoPeDPP for live cell imaging, mitophagy or singlet Oxygen (1O2) detection…). Here I’d like to introduce 3 new molecules developed by Focus Biomolecules to further decipher mitochondrial biology.

#1 – Mito-Photo-DNP | A photo-activated protophore

Mito-Photo-DNP is a mitochondria specific photo-activated protonophore (cat. nr 21910-1580).

Mito-Photo-DNP (cat. nr 10-1580-0001). Source: Focus Biomolecules tebu-bio

Molecular structure of the Mito-Photo-DNP. Source: Focus Biomolecules.

This molecule consists of three structural units composed by (1) a caged  mitochondrial uncoupler (protonophore), (2) a photocleavable linker coupled to (3) a triphenylphosphonium moiety. (Chalmers et al. (2012), J Am Chem Soc, DOI: 10.1021/ja2077922)

 

dyrect-jc-1-mito-health-assay-kit-cat-nr-233m1890-source-markergene-tebu-bio

Dyrect JC-1 Mito Health Assay Kit cat. nr 233M1890. Source: MarkerGene tebu-bio.

The triphenylphosphonium moiety unables the delivery of molecules of interest to mitochondria without the need to alter genes or protein structure. In the case of Mito-Photo-DNP, the molecule delivered is a UV-induced proton transporter (a.k.a. protonophore).

 

After incubation with Mito-Photo-DNP at 200 nM, the cells of interest can be irradiated with UV light at 355 nm. The UV treatment leads to a temporal control of the mitochondrial functions. Mito-Photo-DNP can be thus employed for selective uncoupling of either individual or a small number of mitochondria within a cell when used with mitochondrial membrane potential-sensitive fluorescence imaging dyes like JC-1MitoView Orange or MitoView Red dyes.

# 2 – M-1 | A Mitochondrial Fusion Promoter

M-1 is a cell permeable promoter of mitochondrial Fusion (cat. nr 21910-1598). Derived from phenylhydrazone, this compound enhances mitochondrial fusion and protects cells from mitochondrial fragmentation seen with cell death. Interestingly, M-1 does not interfere with endoplasmic reticulum  & lysosome morphology.

These events can be analyzed with immuno-assays by using, for example, primary antibodies specific for Mitofusin1 (anti-MFN1 [clone 11E9-1H12]), or Mitofusin-2 (anti-MFN2 [clone 4H8]).

icc-staining-of-hepg2-cells-with-mfn1-monoclonal-antibody-clone-11e9-1h12-cat-nr-157mab10282-with-fitc-green-source-abnova-tebu-bio

Immunocytochemical staining of HepG2 cells with Mfn1 monoclonal antibody, clone 11E9-1H12 (cat. nr 157MAB10282, green). Nuclei and actin were counterstained with Dapi (blue) and Phalloidin (red). Source: Abnova | tebu-bio.

immunoperoxidase-of-monoclonal-antibody-to-mfn2-clone-4h8-cat-nr-157h00009927-m03-on-formalin-fixed-paraffin-embedded-human-heart-source-abnova-tebu-bio

Immunoperoxidase of monoclonal antibody to MFN2 , clone 4H8 (cat. nr 157H00009927-M03) on formalin-fixed paraffin-embedded human heart . Source: Abnova | tebu-bio.

 

#3- Cereulide | An inducer of mitochondrial swelling

Acting as an ionophore with a high affinity to potassium cations, Cereulide induces mitochondrial swelling (cat. nr 21910-2746).

mitopt-cat-nr-233m0845-kit-allows-the-easy-distinction-between-non-apoptotic-red-fluorescent-cells-and-apoptotic-green-fluorescent-cells

Distinction between non-apoptotic red fluorescent cells and apoptotic green fluorescent cells with the MitoPT™ kit (cat. nr 233M0845).

Cereulide is a potent cytotoxin produced by Bacillus cereus that destroys mitochondria. This molecule leads to the loss of the membrane potential and uncoupling of oxidative phosphorylation in the mitochondria.

The detection of the mitochondrial permeability transition event provides an early indication of the initiation of cellular apoptosis that can be measured by the change in the membrane potential across the mitochondrial membrane. The MitoPT™ kit allows the easy distinction between non-apoptotic red fluorescent cells and apoptotic green fluorescent cells.

7 additional molecules most used by our customers for studying mitochondrial biology

Molecule Function
FCCP Uncoupler of oxidative phosphorylation
Mdivi-1 Mitochondrial division inhibitor
MitoBlock-6 Mitochondrial import inhibitor
Mitoquinone Mitochondria-targeted antioxidant
Mito-TEMPO Mitochondria-targeted antioxidant
Oligomycin Mitochondrial ATPase inhibitor
Valinomycin Induces loss of mitochondrial membrane potential

Thank you to the Focus Biomolecules team for their support in selecting these mitochondrial-related molecules among their comprehensive offer of compounds.

 

Subscribe to e-Newsletters on your favourite topicsLearn more about tools like this by subscribing to thematic newsletters on your favourite research topics 

Are you looking for a particular small molecule for studying mitochondrial functions?

Leave your questions below!

Philippe Fixe, PhD
Written by Philippe Fixe, PhD
Philippe Fixe is Marketing Director at tebu-bio, passionate about innovation and R&D in Life sciences, Biotechnology, Medical research, Drug discovery, and also a keen photographer.