MilliporeSigma
  • The Unique Cysteine of F-ATP Synthase OSCP Subunit Participates in Modulation of the Permeability Transition Pore.

The Unique Cysteine of F-ATP Synthase OSCP Subunit Participates in Modulation of the Permeability Transition Pore.

Cell reports (2020-09-03)
Michela Carraro, Kristen Jones, Geppo Sartori, Marco Schiavone, Salvatore Antonucci, Roza Kucharczyk, Jean-Paul di Rago, Cinzia Franchin, Giorgio Arrigoni, Michael Forte, Paolo Bernardi
ABSTRACT

The mitochondrial permeability transition pore (PTP) is a Ca2+-activated channel that plays a key role in cell death. Thiol oxidation facilitates PTP opening, yet the targets and molecular mechanisms still await a definition. Here, we investigate the role of C141 of F-ATP synthase oligomycin sensitivity conferral protein (OSCP) subunit in PTP modulation by oxidation. We find that the OSCP C141S mutation confers resistance to PTP opening and cell death by diamide and MitoParaquat only when cyclophilin D (CyPD) has been ablated, a protective role that can be explained by CyPD shielding C141 from oxidants. The mutation decreases apoptosis in zebrafish embryos, indicating that this OSCP residue is involved in development. Site-directed mutagenesis in yeast suggests that other conserved cysteines in the α, γ, and c subunits of F-ATP synthase are not involved in PTP modulation. Thus, OSCP provides a strategic site that regulates PTP opening by the interplay between CyPD (un)binding and thiol oxidation-reduction.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Iodoacetamide, ≥99% (NMR), crystalline
Supelco
Calcium ionophore II, Selectophore, function tested
Sigma-Aldrich
Oligomycin, A mixture of A, B, and C isomers.
Sigma-Aldrich
Dulbecco′s Modified Eagle′s Medium, Without glucose, L-glutamine, phenol red, sodium pyruvate and sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone, ≥98% (TLC), powder
Sigma-Aldrich
Monoclonal Anti-Vinculin antibody produced in mouse, clone VIN-11-5, ascites fluid
Sigma-Aldrich
Ionomycin calcium salt from Streptomyces conglobatus, powder, ≥98% (HPLC)
Sigma-Aldrich
L-Lactic Dehydrogenase from bovine heart, Type III, ammonium sulfate suspension, ≥500 units/mg protein
Roche
NADH, Grade II, disodium salt
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
Antimycin A from Streptomyces sp.
Sigma-Aldrich
Rotenone, ≥95%
Sigma-Aldrich
Cyclosporin A, from Tolypocladium inflatum, ≥95% (HPLC), solid
Sigma-Aldrich
Phospho(enol)pyruvic acid trisodium salt hydrate, ≥97% (enzymatic)
Sigma-Aldrich
Digitonin, Used as non-ionic detergent
Sigma-Aldrich
Phenylarsine oxide, ≥97%, powder
Sigma-Aldrich
Diamide
Sigma-Aldrich
Alamethicin from Trichoderma viride, ≥98% (HPLC)