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).
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)
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-1, MitoView 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]).
#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).
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.
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Since the release of this post, a novel small molecule derived from cellular slime mold Polysphondylium pseudo-candidum has been released – Ppc-1 MitoUncoupler: http://www.tebu-bio.com/Product/194FDV-0011/Ppc_1_MitoUncoupler.html
Ppc-1 MitoUncoupler enhances oxygen consumption in a dose-dependent manner and acts as moderate mitochondrial oxidative phosphorylation uncoupler without adverse effects on ATP production. A post will be released soon regarding this new molecule from Funakoshi. So stay tuned!