Reconstituted adenine nucleotide translocase forms a channel for small molecules comparable to the mitochondrial permeability transition pore

FEBS Letters ◽  
1998 ◽  
Vol 426 (1) ◽  
pp. 97-101 ◽  
Author(s):  
Alexander Rück ◽  
Max Dolder ◽  
Theo Wallimann ◽  
Dieter Brdiczka
Keyword(s):  
Small Molecules ◽  
Mitochondrial Permeability Transition Pore ◽  
Mitochondrial Permeability Transition ◽  
Adenine Nucleotide ◽  
Permeability Transition Pore ◽  
Permeability Transition ◽  
Adenine Nucleotide Translocase ◽  
Mitochondrial Permeability

scholarly journals Role of critical thiol groups on the matrix surface of the adenine nucleotide translocase in the mechanism of the mitochondrial permeability transition pore

2002 ◽  
Vol 367 (2) ◽  
pp. 541-548 ◽  
Author(s):  
Gavin P. McSTAY ◽  
Samantha J. CLARKE ◽  
Andrew P. HALESTRAP
Keyword(s):  
Mitochondrial Permeability Transition Pore ◽  
Mitochondrial Permeability Transition ◽  
Adenine Nucleotide ◽  
Permeability Transition Pore ◽  
Permeability Transition ◽  
Adenine Nucleotide Translocase ◽  
Cross Linking ◽  
Thiol Groups ◽  
Mitochondrial Permeability ◽  
Pre Treatment

Opening of the mitochondrial permeability transition pore (MPTP) is sensitized to [Ca2+] by oxidative stress (diamide) and phenylarsine oxide (PAO). We have proposed that both agents cross-link two thiol groups on the adenine nucleotide translocase (ANT) involved in ADP and cyclophilin-D (CyP-D) binding. Here, we demonstrate that blocking Cys160 with 80μM eosin 5-maleimide (EMA) or 500μM N-ethylmaleimide (NEM) greatly decreased ADP inhibition of the MPTP. The ability of diamide, but not PAO, to block ADP inhibition of the MPTP was antagonized by treatment of mitochondria with 50μM NEM to alkylate matrix glutathione. Binding of detergent-solubilized ANT to a PAO-affinity matrix was prevented by pre-treatment of mitochondria with diamide, EMA or PAO, but not NEM. EMA binding to the ANT in submitochondrial particles (SMPs) was prevented by pre-treatment of mitochondria with either PAO or diamide, implying that both agents modify Cys160. Diamide and PAO pre-treatments also inhibited binding of solubilized ANT to a glutathione S-transferase—CyP-D affinity column, both effects being blocked by 100μM EMA. Intermolecular cross-linking of adjacent ANT molecules via Cys57 by copper phenanthroline treatment of SMPs was abolished by pre-treatment of mitochondria with diamide and PAO, but not with EMA. Our data suggest that PAO and diamide cause intramolecular cross-linking between Cys160 and Cys257 directly (not antagonized by 50μM NEM) or using glutathione (antagonized by 50μM NEM) respectively. This cross-linking stabilizes the ‘c’ conformation of the ANT, reducing the reactivity of Cys57, while enhancing CyP-D binding to the ANT and antagonizing ADP binding. The two effects together greatly sensitize the MPTP to [Ca2+].

scholarly journals The mitochondrial permeability transition pore and its role in cell death

1999 ◽  
Vol 341 (2) ◽  
pp. 233-249 ◽  
Author(s):  
Martin CROMPTON
Keyword(s):  
Cell Death ◽  
Mitochondrial Permeability Transition Pore ◽  
Mitochondrial Permeability Transition ◽  
Adenine Nucleotide ◽  
Permeability Transition Pore ◽  
Permeability Transition ◽  
Adenine Nucleotide Translocase ◽  
Mitochondrial Permeability ◽  
Ischaemia Reperfusion ◽  
Pore Opening

This article reviews the involvement of the mitochondrial permeability transition pore in necrotic and apoptotic cell death. The pore is formed from a complex of the voltage-dependent anion channel (VDAC), the adenine nucleotide translocase and cyclophilin-D (CyP-D) at contact sites between the mitochondrial outer and inner membranes. In vitro, under pseudopathological conditions of oxidative stress, relatively high Ca2+ and low ATP, the complex flickers into an open-pore state allowing free diffusion of low-Mr solutes across the inner membrane. These conditions correspond to those that unfold during tissue ischaemia and reperfusion, suggesting that pore opening may be an important factor in the pathogenesis of necrotic cell death following ischaemia/reperfusion. Evidence that the pore does open during ischaemia/reperfusion is discussed. There are also strong indications that the VDAC-adenine nucleotide translocase-CyP-D complex can recruit a number of other proteins, including Bax, and that the complex is utilized in some capacity during apoptosis. The apoptotic pathway is amplified by the release of apoptogenic proteins from the mitochondrial intermembrane space, including cytochrome c, apoptosis-inducing factor and some procaspases. Current evidence that the pore complex is involved in outer-membrane rupture and release of these proteins during programmed cell death is reviewed, along with indications that transient pore opening may provoke ‘accidental’ apoptosis.

scholarly journals Inhibition of mitochondrial permeability transition by deletion of the ANT family and CypD

2018 ◽  
Author(s):  
Jason Karch ◽  
Michael J. Bround ◽  
Hadi Khalil ◽  
Michelle A. Sargent ◽  
Nadina Latchman ◽  
...  
Keyword(s):  
Mitochondrial Permeability Transition Pore ◽  
Mitochondrial Permeability Transition ◽  
Adenine Nucleotide ◽  
Permeability Transition Pore ◽  
Permeability Transition ◽  
Liver Mitochondria ◽  
Null Mouse ◽  
Inner Membrane ◽  
Membrane Pore ◽  
Mitochondrial Permeability

AbstractThe mitochondrial permeability transition pore (MPTP) has resisted molecular identification for decades. The original model of the MPTP had the adenine nucleotide translocator (ANT) as the inner membrane pore-forming component. Indeed, reconstitution experiments showed that recombinant or purified ANT generates MPTP-like pores in lipid bilayers. This model was challenged when mitochondria from Ant1/2 double null mouse liver still showed MPTP activity. Because mice contain and express 3 Ant genes, here we reinvestigated the genetic basis for the ANTs as comprising the MPTP. Liver mitochondria from Ant1, Ant2, and Ant4 deficient mice were highly refractory to Ca2+-induced MPT, and when also given cyclosporine A, MPT was completely inhibited. Moreover, liver mitochondria from mice with quadruple deletion of Ant1, Ant2, Ant4 and Ppif (cyclophilin D, target of CsA) lacked Ca2+-induced MPT. Finally, inner membrane patch clamping in mitochondria from Ant1, Ant2 and Ant4 triple null mouse embryonic fibroblasts (MEFs) showed a loss of MPT-like pores. Our findings suggest a new model of MPT consisting of two distinct molecular components, one of which is the ANTs and the other of which is unknown but requires CypD.One Sentence SummaryGenetic deletion of Ant1/2/4 and Ppif in mice fully inhibits the mitochondrial permeability transition pore

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