title: Titration of lysine residues on adenine nucleotide translocase by fluorescamine induces permeability transition.
abstract: Chemical modification of primary amino groups of mitochondrial membrane proteins by the fluorescent probe fluorescamine induces non-specific membrane permeabilisation. Titration of the lysine ϵ-amino group promoted efflux of accumulated Ca(2+), collapse of transmembrane potential and mitochondrial swelling. Ca(2+) release was inhibited by cyclosporin A. Considering the latter, we assumed that fluorescamine induces permeability transition. Carboxyatractyloside also inhibited the reaction. Using a polyclonal antibody for adenine nucleotide translocase, Western blot analysis showed that the carrier appeared labelled with the fluorescent probe. The results point out the importance of the ϵ-amino group of lysine residues, located in the adenine nucleotide carrier, on the modulation of membrane permeability, since its blockage suffices to promote opening of the non-specific nanopore.
authors.affiliation: Departamento de Bioquímica Instituto Nacional de Cardiología Ignacio Chávez, Mexico, D.F., 014080, Mexico.
authors.forename: Mabel
authors.initials: M
authors.lastname: Buelna-Chontal
authors.forename: Natalia
authors.initials: N
authors.lastname: Pavón
authors.forename: Francisco
authors.initials: F
authors.lastname: Correa
authors.forename: Luz
authors.initials: L
authors.lastname: Hernández-Esquivel
authors.forename: Edmundo
authors.initials: E
authors.lastname: Chávez
mesh_terms: D000818:Animals; D001278:Atractyloside / Q000031:analogs & derivatives / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects* / Q000502:physiology; D005450:Fluorescamine / Q000494:pharmacology*; D017136:Ion Transport / Q000187:drug effects / Q000502:physiology; D008239:Lysine / Q000378:metabolism*; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects* / Q000502:physiology; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D000226:Mitochondrial ADP, ATP Translocases / Q000187:drug effects / Q000378:metabolism*; D008933:Mitochondrial Swelling / Q000187:drug effects / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar
chemical_list: D001278:Atractyloside; D005450:Fluorescamine; D000226:Mitochondrial ADP, ATP Translocases; D008239:Lysine; C003853:carboxyatractyloside; D002118:Calcium
keywords: adenine nucleotide carrier; calcium; fluorescamine; mitochondria; permeability transition