Mitochondrial targeting of cyclosporin A enables selective inhibition of cyclophilin-D and enhanced cytoprotection after glucose and oxygen deprivation
| Autor: | David L. Selwood, Sylvanie Malouitre, Martin Crompton, Henry Dube |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2009 |
| Předmět: |
HBSS
Hanks balanced salt solution CyP-D+ cell line overexpressing CyP-D Excitotoxicity Mitochondria Liver Mitochondrion medicine.disease_cause Hippocampus Biochemistry necrosis Rats Sprague-Dawley Cyclophilins ESI–MS electrospray ionization MS 0302 clinical medicine Fmoc fluoren-9-ylmethoxycarbonyl cyclosporin A (CsA) CsA cyclosporin A Cyclosporin a PT permeability transition NBQX 2 3-dihydro-6-nitro-7-sulfamoylbenzoquinoxaline Enzyme Inhibitors Inner mitochondrial membrane Cells Cultured Membrane Potential Mitochondrial Neurons 0303 health sciences Molecular Structure TMRE tetramethylrhodamine ethyl ester Calcineurin DMEM Dulbecco's minimal essential medium Glutamate receptor L-NAME NG-nitro-L-arginine-methyl ester NBA Neurobasal A Cytoprotection DCM dichloromethane Cell biology OGD oxygen and glucose deprivation LDA lithium diisopropylamide hippocampal neuron CyP cyclophilin Cyclosporine SMBz-CsA [sarcosine-3(4-methylbenzoate)]-CsA ischaemia DMF dimethylformamide Cyclophilin A mtCsA mitochondrially targeted CsA THF tetrahydrofuran Cyclophilin D Research Article Protein Binding Programmed cell death Cell Survival Glutamic Acid Biology glutamate toxicity 03 medical and health sciences PPIase peptidylprolyl cis–trans isomerase FBS fetal bovine serum Cell Line Tumor Cyclophilin (CyP) cyclosporin A (CsA) glutamate toxicity hippocampal neuron ischaemia necrosis medicine PyBOP benzotriazol-1-yl-tris-pyrrolidinophosphonium hexafluorophosphate Animals cyclophilin (CyP) Molecular Biology 030304 developmental biology NMDA N-methyl-D-aspartate I/R ischaemia/reperfusion TPP+ triphenylphosphonium Cell Biology medicine.disease Rats Oxygen Glucose Animals Newborn Reperfusion injury 030217 neurology & neurosurgery |
| Zdroj: | Biochemical Journal, 425 (1) pp. 137-148. (2009) Biochemical Journal |
| Popis: | CsA (cyclosporin A) is a hydrophobic undecapeptide that inhibits CyPs (cyclophilins), a family of PPIases (peptidylprolyl cis–trans isomerases). In some experimental models, CsA offers partial protection against lethal cell injury brought about by transient ischaemia; this is believed to reflect inhibition of CyP-D, a mitochondrial isoform that facilitates formation of the permeability transition pore in the mitochondrial inner membrane. To evaluate this further, we have targeted CsA to mitochondria so that it becomes selective for CyP-D in cells. This was achieved by conjugating the inhibitor to the lipophilic triphenylphosphonium cation, enabling its accumulation in mitochondria due to the inner membrane potential. In a cell-free system and in B50 neuroblastoma cells the novel reagent (but not CsA itself) preferentially inhibited CyP-D over extramitochondrial CyP-A. In hippocampal neurons, mitochondrial targeting markedly enhanced the capacity of CsA to prevent cell necrosis brought about by oxygen and glucose deprivation, but largely abolished its capacity to inhibit glutamate-induced cell death. It is concluded that CyP-D has a major pathogenic role in ‘energy failure’, but not in glutamate excitotoxicity, where cytoprotection primarily reflects CsA interaction with extramitochondrial CyPs and calcineurin. Moreover, the therapeutic potential of CsA against ischaemia/reperfusion injuries not involving glutamate may be improved by mitochondrial targeting. |
| Databáze: | OpenAIRE |
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