E S5 Properties of Nif genes in Groups III and IV.Author ContributionsConceived and created the experiments: JBH KJK DCR ANG. Analyzed the information: JBH KJK DCR ANG. Wrote the paper: JBH KJK DCR ANG.(PDF)
THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 289, NO. 29, pp. 203770385, July 18, 2014 2014 by The American Society for Biochemistry and Molecular Biology, Inc. Published inside the U.S.A.NCLX Protein, but Not LETM1, Mediates Mitochondrial Ca2 Extrusion, Thereby Limiting Ca2 -induced NAD(P)H Production and Modulating Matrix Redox State*Received for publication, December 18, 2013, and in revised kind, Might 12, 2014 Published, JBC Papers in Press, June four, 2014, DOI 10.1074/jbc.M113.Umberto De Marchi, Jaime Santo-Domingo Cyril Castelbou Israel Sekler Andreas Wiederkehr, and Nicolas Demaurex In the Mitochondrial Function, Nestle Institute of Overall health Sciences, EPFL Innovation Park, Constructing G, CH-1015 Lausanne, Switzerland, the �Department of Cell Physiology and Metabolism, University of Geneva, Rue Michel-Servet, 1, CH-1211 Geneve, ` Switzerland, and also the epartment of Physiology, Ben-Gurion University of Negev, Beer-Sheva 84105, IsraelBackground: Irrespective of whether mitochondrial Ca2 extrusion is mediated by NCLX (mitochondrial sodium/calcium exchanger) or LETM1 (leucine zipper-EF-hand-containing transmembrane protein 1) and controls matrix redox state is unknown. Results: NCLX, but not LETM1, increases Ca2 extrusion, limits NAD(P)H production, and promotes matrix oxidation. Conclusion: NCLX controls the duration of matrix Ca2 elevations and their influence on redox signaling. Significance: NCLX is usually a prospective target for the therapy of redox-dependent ailments. Mitochondria capture and subsequently release Ca2 ions, thereby sensing and shaping cellular Ca2 signals. The Ca2 uniporter MCU mediates Ca2 uptake, whereas NCLX (mitochondrial Na/Ca exchanger) and LETM1 (leucine zipper-EFhand-containing transmembrane protein 1) have been proposed to exchange Ca2 against Na or H , respectively. Right here we study the function of those ion exchangers in mitochondrial Ca2 extrusion and in Ca2 -metabolic coupling. Each NCLX and LETM1 proteins were expressed in HeLa cells mitochondria. The rate of mitochondrial Ca2 efflux, measured using a genetically encoded indicator during agonist stimulations, improved with all the amplitude of mitochondrial Ca2 ([Ca2 ]mt) elevations. NCLX overexpression enhanced the prices of Ca2 efflux, whereas growing LETM1 levels had no effect on Ca2 extrusion. The fluorescence of your redox-sensitive probe roGFP improved in the course of [Ca2 ]mt elevations, indicating a net reduction of the matrix.3-Hydroxydodecanoic acid In Vivo This redox response was abolished by NCLX overexpression and restored by the Na /Ca2 exchanger inhibitor CGP37157.Dp44mT Protocol The [Ca2 ]mt elevations had been related with increases within the autofluorescence of NAD(P)H, whose amplitude was strongly lowered by NCLX overexpression, an impact reverted by Na / Ca2 exchange inhibition.PMID:24182988 We conclude that NCLX, but not LETM1, mediates Ca2 extrusion from mitochondria. By controlling the duration of matrix Ca2 elevations, NCLX contributes for the regulation of NAD(P)H production and to the conversion of Ca2 signals into redox changes.* This operate was supported by Swiss National Foundation Grant 310030B_133126 (to N. D.). To whom correspondence could be addressed: Mitochondrial Function, Nestle Institute of Overall health Sciences, EPFL Innovation Park, Bldg. G, CH-1015 Lausanne, Switzerland. Tel.: 41-21-632-6122; E-mail: Umberto.DeMarchi@ rd.nestle. 2 To whom correspon.
