title: Cystathionine β-synthase regulates endothelial function via protein S-sulfhydration.
abstract: Deficiencies of the human cystathionine β-synthase (CBS) enzyme are characterized by a plethora of vascular disorders and hyperhomocysteinemia. However, several clinical trials demonstrated that despite reduction in homocysteine levels, disease outcome remained unaffected, thus the mechanism of endothelial dysfunction is poorly defined. Here, we show that the loss of CBS function in endothelial cells (ECs) leads to a significant down-regulation of cellular hydrogen sulfide (H2S) by 50% and of glutathione (GSH) by 40%. Silencing CBS in ECs compromised phenotypic and signaling responses to the VEGF that were potentiated by decreased transcription of VEGF receptor (VEGFR)-2 and neuropilin (NRP)-1, the primary receptors regulating endothelial function. Transcriptional down-regulation of VEGFR-2 and NRP-1 was mediated by a lack in stability of the transcription factor specificity protein 1 (Sp1), which is a sulfhydration target of H2S at residues Cys68 and Cys755. Reinstating H2S but not GSH in CBS-silenced ECs restored Sp1 levels and its binding to the VEGFR-2 promoter and VEGFR-2, NRP-1 expression, VEGF-dependent proliferation, and migration phenotypes. Thus, our study emphasizes the importance of CBS-mediated protein S-sulfhydration in maintaining vascular health and function.-Saha, S., Chakraborty, P. K., Xiong, X., Dwivedi, S. K. D., Mustafi, S. B., Leigh, N. R., Ramchandran, R., Mukherjee, P., Bhattacharya, R. Cystathionine β-synthase regulates endothelial function via protein S-sulfhydration.
authors.affiliation: *Peggy and Charles Stephenson Cancer Center, Department of Obstetrics and Gynecology, Department of Pathology, and Department of Cell Biology, University of Oklahoma Health Science Center, Oklahoma City, Oklahoma, USA; and Developmental Vascular Biology Program and Zebrafish Drug Screening Core, Department of Obstetrics and Gynecology, The Medical College of Wisconsin, Milwaukee, Wisconsin, USA.
authors.forename: Sounik
authors.initials: S
authors.lastname: Saha
authors.affiliation: *Peggy and Charles Stephenson Cancer Center, Department of Obstetrics and Gynecology, Department of Pathology, and Department of Cell Biology, University of Oklahoma Health Science Center, Oklahoma City, Oklahoma, USA; and Developmental Vascular Biology Program and Zebrafish Drug Screening Core, Department of Obstetrics and Gynecology, The Medical College of Wisconsin, Milwaukee, Wisconsin, USA.
authors.forename: Prabir K
authors.initials: PK
authors.lastname: Chakraborty
authors.affiliation: *Peggy and Charles Stephenson Cancer Center, Department of Obstetrics and Gynecology, Department of Pathology, and Department of Cell Biology, University of Oklahoma Health Science Center, Oklahoma City, Oklahoma, USA; and Developmental Vascular Biology Program and Zebrafish Drug Screening Core, Department of Obstetrics and Gynecology, The Medical College of Wisconsin, Milwaukee, Wisconsin, USA.
authors.forename: Xunhao
authors.initials: X
authors.lastname: Xiong
authors.affiliation: *Peggy and Charles Stephenson Cancer Center, Department of Obstetrics and Gynecology, Department of Pathology, and Department of Cell Biology, University of Oklahoma Health Science Center, Oklahoma City, Oklahoma, USA; and Developmental Vascular Biology Program and Zebrafish Drug Screening Core, Department of Obstetrics and Gynecology, The Medical College of Wisconsin, Milwaukee, Wisconsin, USA.
authors.forename: Shailendra Kumar Dhar
authors.initials: SK
authors.lastname: Dwivedi
authors.affiliation: *Peggy and Charles Stephenson Cancer Center, Department of Obstetrics and Gynecology, Department of Pathology, and Department of Cell Biology, University of Oklahoma Health Science Center, Oklahoma City, Oklahoma, USA; and Developmental Vascular Biology Program and Zebrafish Drug Screening Core, Department of Obstetrics and Gynecology, The Medical College of Wisconsin, Milwaukee, Wisconsin, USA.
authors.forename: Soumyajit Banerjee
authors.initials: SB
authors.lastname: Mustafi
authors.affiliation: *Peggy and Charles Stephenson Cancer Center, Department of Obstetrics and Gynecology, Department of Pathology, and Department of Cell Biology, University of Oklahoma Health Science Center, Oklahoma City, Oklahoma, USA; and Developmental Vascular Biology Program and Zebrafish Drug Screening Core, Department of Obstetrics and Gynecology, The Medical College of Wisconsin, Milwaukee, Wisconsin, USA.
authors.forename: Noah R
authors.initials: NR
authors.lastname: Leigh
authors.affiliation: *Peggy and Charles Stephenson Cancer Center, Department of Obstetrics and Gynecology, Department of Pathology, and Department of Cell Biology, University of Oklahoma Health Science Center, Oklahoma City, Oklahoma, USA; and Developmental Vascular Biology Program and Zebrafish Drug Screening Core, Department of Obstetrics and Gynecology, The Medical College of Wisconsin, Milwaukee, Wisconsin, USA.
authors.forename: Ramani
authors.initials: R
authors.lastname: Ramchandran
authors.affiliation: *Peggy and Charles Stephenson Cancer Center, Department of Obstetrics and Gynecology, Department of Pathology, and Department of Cell Biology, University of Oklahoma Health Science Center, Oklahoma City, Oklahoma, USA; and Developmental Vascular Biology Program and Zebrafish Drug Screening Core, Department of Obstetrics and Gynecology, The Medical College of Wisconsin, Milwaukee, Wisconsin, USA priyabrata-mukherjee@ouhsc.edu resham-bhattacharya@ouhsc.edu.
authors.forename: Priyabrata
authors.initials: P
authors.lastname: Mukherjee
authors.affiliation: *Peggy and Charles Stephenson Cancer Center, Department of Obstetrics and Gynecology, Department of Pathology, and Department of Cell Biology, University of Oklahoma Health Science Center, Oklahoma City, Oklahoma, USA; and Developmental Vascular Biology Program and Zebrafish Drug Screening Core, Department of Obstetrics and Gynecology, The Medical College of Wisconsin, Milwaukee, Wisconsin, USA priyabrata-mukherjee@ouhsc.edu resham-bhattacharya@ouhsc.edu.
authors.forename: Resham
authors.initials: R
authors.lastname: Bhattacharya
mesh_terms: D002465:Cell Movement; D049109:Cell Proliferation; D003541:Cystathionine beta-Synthase / Q000235:genetics / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D005978:Glutathione / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D006801:Humans; D006862:Hydrogen Sulfide / Q000378:metabolism*; D039921:Neuropilins / Q000235:genetics / Q000378:metabolism; D015290:Second Messenger Systems; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism
chemical_list: D039921:Neuropilins; D042461:Vascular Endothelial Growth Factor A; C000588866:KDR protein, human; D040301:Vascular Endothelial Growth Factor Receptor-2; D003541:Cystathionine beta-Synthase; D005978:Glutathione; D006862:Hydrogen Sulfide
keywords: VEGFRs; angiogenesis; metabolism; signal transduction