title: Improved tag-switch method reveals that thioredoxin acts as depersulfidase and controls the intracellular levels of protein persulfidation.
abstract: Hydrogen sulfide (H2S) has emerged as a signalling molecule capable of regulating several important physiological functions such as blood pressure, neurotransmission and inflammation. The mechanisms behind these effects are still largely elusive and oxidative posttranslational modification of cysteine residues (protein persulfidation orS-sulfhydration) has been proposed as the main pathway for H2S-induced biological and pharmacological effects. As a signalling mechanism, persulfidation has to be controlled. Using an improved tag-switch assay for persulfide detection we show here that protein persulfide levels are controlled by the thioredoxin system. Recombinant thioredoxin showed an almost 10-fold higher reactivity towards cysteine persulfide than towards cystine and readily cleaved protein persulfides as well. This reaction resulted in H2S release suggesting that thioredoxin could be an important regulator of H2S levels from persulfide pools. Inhibition of the thioredoxin system caused an increase in intracellular persulfides, highlighting thioredoxin as a major protein depersulfidase that controls H2S signalling. Finally, using plasma from HIV-1 patients that have higher circulatory levels of thioredoxin, we could prove depersulfidase rolein vivo.
authors.affiliation: Department of Chemistry and Pharmacy , Friedrich-Alexander University of Erlangen-Nuremberg , Erlangen , Germany . Email: milos.filipovic@ibgc.cnrs.fr.
authors.forename: Rudolf
authors.initials: R
authors.lastname: Wedmann
authors.affiliation: Department of Chemistry and Pharmacy , Friedrich-Alexander University of Erlangen-Nuremberg , Erlangen , Germany . Email: milos.filipovic@ibgc.cnrs.fr.
authors.forename: Constantin
authors.initials: C
authors.lastname: Onderka
authors.affiliation: Department of Chemistry and Pharmacy , Friedrich-Alexander University of Erlangen-Nuremberg , Erlangen , Germany . Email: milos.filipovic@ibgc.cnrs.fr.
authors.forename: Shengwei
authors.initials: S
authors.lastname: Wei
authors.affiliation: Charité Campus Virchow , Nephrology/Intensive Care , Berlin , Germany.
authors.forename: István András
authors.initials: IA
authors.lastname: Szijártó
authors.affiliation: Department of Chemistry and Pharmacy , Friedrich-Alexander University of Erlangen-Nuremberg , Erlangen , Germany . Email: milos.filipovic@ibgc.cnrs.fr.
authors.forename: Jan Lj
authors.initials: JL
authors.lastname: Miljkovic
authors.affiliation: Department of Chemistry , University of Belgrade , Belgrade , Serbia.
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Mitrovic
authors.affiliation: Department of Chemistry and Pharmacy , Friedrich-Alexander University of Erlangen-Nuremberg , Erlangen , Germany . Email: milos.filipovic@ibgc.cnrs.fr.
authors.forename: Mike
authors.initials: M
authors.lastname: Lange
authors.affiliation: Department of Chemistry and Pharmacy , Friedrich-Alexander University of Erlangen-Nuremberg , Erlangen , Germany . Email: milos.filipovic@ibgc.cnrs.fr.
authors.forename: Sergey
authors.initials: S
authors.lastname: Savitsky
authors.affiliation: Department of Biological Chemistry , University of Michigan , Ann Arbor , USA.
authors.forename: Pramod Kumar
authors.initials: PK
authors.lastname: Yadav
authors.affiliation: University of Exeter Medical School , St. Luke's Campus , Exeter , UK ; Biosciences , College of Life and Environmental Sciences of Biosciences , University of Exeter , Streatham Campus , Exeter , Devon , UK.
authors.forename: Roberta
authors.initials: R
authors.lastname: Torregrossa
authors.affiliation: Infectious Diseases Section , Department of Internal Medicine 3 , Universitätsklinikum Erlangen , Friedrich-Alexander-University, Erlangen-Nürnberg , Germany.
authors.forename: Ellen G
authors.initials: EG
authors.lastname: Harrer
authors.affiliation: Infectious Diseases Section , Department of Internal Medicine 3 , Universitätsklinikum Erlangen , Friedrich-Alexander-University, Erlangen-Nürnberg , Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Harrer
authors.affiliation: Department of Biochemistry , Graduate School of Pharmaceutical Sciences , Keio University , Tokyo , Japan.
authors.forename: Isao
authors.initials: I
authors.lastname: Ishii
authors.affiliation: Charité Campus Virchow , Nephrology/Intensive Care , Berlin , Germany.
authors.forename: Maik
authors.initials: M
authors.lastname: Gollasch
authors.affiliation: Biosciences , College of Life and Environmental Sciences of Biosciences , University of Exeter , Streatham Campus , Exeter , Devon , UK.
authors.forename: Mark E
authors.initials: ME
authors.lastname: Wood
authors.affiliation: UMR CNRS 8601 , Université Paris Descartes , Sorbonne Paris Cité , Paris , France.
authors.forename: Erwan
authors.initials: E
authors.lastname: Galardon
authors.affiliation: Department of Chemistry , Washington State University , Pullman , USA.
authors.forename: Ming
authors.initials: M
authors.lastname: Xian
authors.affiliation: University of Exeter Medical School , St. Luke's Campus , Exeter , UK.
authors.forename: Matthew
authors.initials: M
authors.lastname: Whiteman
authors.affiliation: Department of Biological Chemistry , University of Michigan , Ann Arbor , USA.
authors.forename: Ruma
authors.initials: R
authors.lastname: Banerjee
authors.affiliation: Department of Chemistry and Pharmacy , Friedrich-Alexander University of Erlangen-Nuremberg , Erlangen , Germany . Email: milos.filipovic@ibgc.cnrs.fr; Université de Bordeaux , IBGC , UMR 5095 , F-33077 Bordeaux , France ; CNRS , IBGC , UMR 5095 , F-33077 Bordeaux , France.
authors.forename: Milos R
authors.initials: MR
authors.lastname: Filipovic