A clickable probe for versatile characterization of S-nitrosothiols
Jenna L. Clements,
Franziska Pohl,
Pandi Muthupandi,
Stephen C. Rogers,
Jack Mao,
Allan Doctor,
Vladimir B. Birman,
Jason M. Held
Affiliations
Jenna L. Clements
Department of Medicine, Washington University Medical School, St. Louis, MO, 63110, USA
Franziska Pohl
Department of Medicine, Washington University Medical School, St. Louis, MO, 63110, USA
Pandi Muthupandi
Department of Chemistry, Washington University in Saint Louis, St. Louis, MO, 63110, USA
Stephen C. Rogers
Department of Pediatrics and Center for Blood Oxygen Transport and Hemostasis, University of Maryland, School of Medicine, Baltimore, MD, 21201, USA
Jack Mao
Department of Medicine, Washington University Medical School, St. Louis, MO, 63110, USA
Allan Doctor
Department of Pediatrics and Center for Blood Oxygen Transport and Hemostasis, University of Maryland, School of Medicine, Baltimore, MD, 21201, USA
Vladimir B. Birman
Department of Chemistry, Washington University in Saint Louis, St. Louis, MO, 63110, USA
Jason M. Held
Department of Medicine, Washington University Medical School, St. Louis, MO, 63110, USA; Siteman Cancer Center, Washington University Medical School, St. Louis, MO, 63110, USA; Department of Anesthesiology, Washington University Medical School, St. Louis, MO, 63110, USA; Corresponding author. Washington University, School of Medicine, Campus Box 8076, 660 South Euclid Avenue, Saint Louis, MO, 63110, USA.
S-nitrosation of cysteine thiols (SNOs), commonly referred to as S-nitrosylation, is a cysteine oxoform that plays an important role in cellular signaling and impacts protein function and stability. Direct labeling of SNOs in cells with the flexibility to perform a wide range of cellular and biochemical assays remains a bottleneck as all SNO-targeted probes to date employ a single analytical modality such as biotin or a specific fluorophore. We therefore developed a clickable, alkyne-containing SNO probe ‘PBZyn’ based on the o-phosphino-benzoyl group warhead that enables multi-modal analysis via click conjugation. We demonstrate the utility of PBZyn to assay SNOs using in situ cellular imaging, protein blotting and affinity purification, as well as mass spectrometry. The flexible PBZyn probe will greatly facilitate investigation into the regulation of SNOs.
