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S-Sulfhydration of ATP synthase by hydrogen sulfide stimulates mitochondrial bioenergetics.
Pharmacol Res. 2016 11; 113(Pt A):116-124.PR

Abstract

Mammalian cells can utilize hydrogen sulfide (H2S) to support mitochondrial respiration. The aim of our study was to explore the potential role of S-sulfhydration (a H2S-induced posttranslational modification, also known as S-persulfidation) of the mitochondrial inner membrane protein ATP synthase (F1F0 ATP synthase/Complex V) in the regulation of mitochondrial bioenergetics. Using a biotin switch assay, we have detected S-sulfhydration of the α subunit (ATP5A1) of ATP synthase in response to exposure to H2S in vitro. The H2S generator compound NaHS induced S-sulfhydration of ATP5A1 in HepG2 and HEK293 cell lysates in a concentration-dependent manner (50-300μM). The activity of immunocaptured mitochondrial ATP synthase enzyme isolated from HepG2 and HEK293 cells was stimulated by NaHS at low concentrations (10-100nM). Site-directed mutagenesis of ATP5A1 in HEK293 cells demonstrated that cysteine residues at positions 244 and 294 are subject to S-sulfhydration. The double mutant ATP synthase protein (C244S/C294S) showed a significantly reduced enzyme activity compared to control and the single-cysteine-mutated recombinant proteins (C244S or C294S). To determine whether endogenous H2S plays a role in the basal S-sulfhydration of ATP synthase in vivo, we compared liver tissues harvested from wild-type mice and mice deficient in cystathionine-gamma-lyase (CSE, one of the three principal mammalian H2S-producing enzymes). Significantly reduced S-sulfhydration of ATP5A1 was observed in liver homogenates of CSE-/- mice, compared to wild-type mice, suggesting a physiological role for CSE-derived endogenous H2S production in the S-sulfhydration of ATP synthase. Various forms of critical illness (including burn injury) upregulate H2S-producing enzymes and stimulate H2S biosynthesis. In liver tissues collected from mice subjected to burn injury, we detected an increased S-sulfhydration of ATP5A1 at the early time points post-burn. At later time points (when systemic H2S levels decrease) S-sulfhydration of ATP5A1 decreased as well. In conclusion, H2S induces S-sulfhydration of ATP5A1 at C244 and C294. This post-translational modification may be a physiological mechanism to maintain ATP synthase in a physiologically activated state, thereby supporting mitochondrial bioenergetics. The sulfhydration of ATP synthase may be a dynamic process, which may be regulated by endogenous H2S levels under various pathophysiological conditions.

Authors+Show Affiliations

Cardiovascular and Metabolic Research Unit, Lakehead University, Thunder Bay, ON, Canada; Department of Surgery, University of Texas Medical Branch, Galveston, TX, USA; Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA.Cardiovascular and Metabolic Research Unit, Lakehead University, Thunder Bay, ON, Canada.Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA.Cardiovascular and Metabolic Research Unit, Lakehead University, Thunder Bay, ON, Canada; Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA.Cardiovascular and Metabolic Research Unit, Lakehead University, Thunder Bay, ON, Canada.School of Human Kinesiology, Laurentian University, Sudbury, ON, Canada.Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA. Electronic address: szabocsaba@aol.com.Cardiovascular and Metabolic Research Unit, Lakehead University, Thunder Bay, ON, Canada; Department of Biology, Laurentian University, ON, Canada. Electronic address: rui.wang@laurentian.ca.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

27553984

Citation

Módis, Katalin, et al. "S-Sulfhydration of ATP Synthase By Hydrogen Sulfide Stimulates Mitochondrial Bioenergetics." Pharmacological Research, vol. 113, no. Pt A, 2016, pp. 116-124.
Módis K, Ju Y, Ahmad A, et al. S-Sulfhydration of ATP synthase by hydrogen sulfide stimulates mitochondrial bioenergetics. Pharmacol Res. 2016;113(Pt A):116-124.
Módis, K., Ju, Y., Ahmad, A., Untereiner, A. A., Altaany, Z., Wu, L., Szabo, C., & Wang, R. (2016). S-Sulfhydration of ATP synthase by hydrogen sulfide stimulates mitochondrial bioenergetics. Pharmacological Research, 113(Pt A), 116-124. https://doi.org/10.1016/j.phrs.2016.08.023
Módis K, et al. S-Sulfhydration of ATP Synthase By Hydrogen Sulfide Stimulates Mitochondrial Bioenergetics. Pharmacol Res. 2016;113(Pt A):116-124. PubMed PMID: 27553984.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - S-Sulfhydration of ATP synthase by hydrogen sulfide stimulates mitochondrial bioenergetics. AU - Módis,Katalin, AU - Ju,YoungJun, AU - Ahmad,Akbar, AU - Untereiner,Ashley A, AU - Altaany,Zaid, AU - Wu,Lingyun, AU - Szabo,Csaba, AU - Wang,Rui, Y1 - 2016/08/20/ PY - 2016/05/23/received PY - 2016/07/12/revised PY - 2016/08/18/accepted PY - 2016/10/21/pubmed PY - 2017/12/21/medline PY - 2016/8/25/entrez KW - ATP synthase KW - Bioenergetics KW - Burn KW - Burn injury KW - Cysteine KW - H(2)S KW - Hydrogen sulfide KW - Mitochondria KW - S-Sulfhydration SP - 116 EP - 124 JF - Pharmacological research JO - Pharmacol Res VL - 113 IS - Pt A N2 - Mammalian cells can utilize hydrogen sulfide (H2S) to support mitochondrial respiration. The aim of our study was to explore the potential role of S-sulfhydration (a H2S-induced posttranslational modification, also known as S-persulfidation) of the mitochondrial inner membrane protein ATP synthase (F1F0 ATP synthase/Complex V) in the regulation of mitochondrial bioenergetics. Using a biotin switch assay, we have detected S-sulfhydration of the α subunit (ATP5A1) of ATP synthase in response to exposure to H2S in vitro. The H2S generator compound NaHS induced S-sulfhydration of ATP5A1 in HepG2 and HEK293 cell lysates in a concentration-dependent manner (50-300μM). The activity of immunocaptured mitochondrial ATP synthase enzyme isolated from HepG2 and HEK293 cells was stimulated by NaHS at low concentrations (10-100nM). Site-directed mutagenesis of ATP5A1 in HEK293 cells demonstrated that cysteine residues at positions 244 and 294 are subject to S-sulfhydration. The double mutant ATP synthase protein (C244S/C294S) showed a significantly reduced enzyme activity compared to control and the single-cysteine-mutated recombinant proteins (C244S or C294S). To determine whether endogenous H2S plays a role in the basal S-sulfhydration of ATP synthase in vivo, we compared liver tissues harvested from wild-type mice and mice deficient in cystathionine-gamma-lyase (CSE, one of the three principal mammalian H2S-producing enzymes). Significantly reduced S-sulfhydration of ATP5A1 was observed in liver homogenates of CSE-/- mice, compared to wild-type mice, suggesting a physiological role for CSE-derived endogenous H2S production in the S-sulfhydration of ATP synthase. Various forms of critical illness (including burn injury) upregulate H2S-producing enzymes and stimulate H2S biosynthesis. In liver tissues collected from mice subjected to burn injury, we detected an increased S-sulfhydration of ATP5A1 at the early time points post-burn. At later time points (when systemic H2S levels decrease) S-sulfhydration of ATP5A1 decreased as well. In conclusion, H2S induces S-sulfhydration of ATP5A1 at C244 and C294. This post-translational modification may be a physiological mechanism to maintain ATP synthase in a physiologically activated state, thereby supporting mitochondrial bioenergetics. The sulfhydration of ATP synthase may be a dynamic process, which may be regulated by endogenous H2S levels under various pathophysiological conditions. SN - 1096-1186 UR - https://www.unboundmedicine.com/medline/citation/27553984/S_Sulfhydration_of_ATP_synthase_by_hydrogen_sulfide_stimulates_mitochondrial_bioenergetics_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1043-6618(16)30482-0 DB - PRIME DP - Unbound Medicine ER -