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Here, we describe a procedure for the identification of S-nitrosothiols that has been used in our laboratory to study the roles of protein S-nitrosylation in the immune responses of Arabidopsis thaliana and other organisms. It employs a modified version of the biotin-switch technique, which we termed the sequential cysteine blocking technique, encompassing the sequential redox-blocking of recombinant proteins followed by LC-MS/MS analysis.
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http://dx.doi.org/10.1007/978-1-4939-3600-7_14 | DOI Listing |
Plant Physiol
December 2024
Department of Biochemistry and Molecular Biology, University of Massachusetts Amherst, Amherst, MA 01003, USA.
Plant fertility is fundamental to plant survival and requires the coordinated interaction of developmental pathways and signaling molecules. Nitric oxide (NO) is a small, gaseous signaling molecule that plays crucial roles in plant fertility as well as other developmental processes and stress responses. NO influences biological processes through S-nitrosation, the posttranslational modification of protein cysteines to S-nitrosocysteine (R-SNO).
View Article and Find Full Text PDFFood Chem
January 2025
Shanghai Frontiers Science Center of Drug Target Identification and Delivery, Engineering Research Center of Cell & Therapeutic Antibody, National Key Laboratory of Innovative Immunotherapy, School of Pharmaceutical Sciences, Shanghai Jiao Tong University, Shanghai 200240, China. Electronic address:
S-nitrosoglutathione (GSNO) is the most important S-nitrosothiol in vivo, which could affect the quality of meat by participating in calcium release, glucose metabolism, proteolysis and apoptosis, therefore may potentially serve as a marker for meat freshness. In this work, a solid-phase extraction (SPE) monolithic spin column modified with gold nanoparticles was prepared for GSNO extraction. The optimized SPE-LC-MS/MS method for GSNO quantification displays low limit of detection (0.
View Article and Find Full Text PDFAdv Sci (Weinh)
March 2024
Department of Molecular Medicine and Neurodegeneration New Medicines Center, The Scripps Research Institute, La Jolla, CA, 92037, USA.
In Alzheimer's disease (AD), dysfunctional mitochondrial metabolism is associated with synaptic loss, the major pathological correlate of cognitive decline. Mechanistic insight for this relationship, however, is still lacking. Here, comparing isogenic wild-type and AD mutant human induced pluripotent stem cell (hiPSC)-derived cerebrocortical neurons (hiN), evidence is found for compromised mitochondrial energy in AD using the Seahorse platform to analyze glycolysis and oxidative phosphorylation (OXPHOS).
View Article and Find Full Text PDFACS Chem Biol
January 2024
Department of Chemistry, Boston College, Chestnut Hill, Massachusetts 02467, United States.
-Nitrosation is a cysteine post-translational modification fundamental to cellular signaling. This modification regulates protein function in numerous biological processes in the nervous, cardiovascular, and immune systems. Small molecule or protein nitrosothiols act as mediators of NO signaling by transferring the NO group (formally NO) to a free thiol on a target protein through a transnitrosation reaction.
View Article and Find Full Text PDFCell Death Dis
April 2023
Redox Biology, Danish Cancer Society Research Center, 2100, Copenhagen, Denmark.
S-nitrosylation is a post-translational modification in which nitric oxide (NO) binds to the thiol group of cysteine, generating an S-nitrosothiol (SNO) adduct. S-nitrosylation has different physiological roles, and its alteration has also been linked to a growing list of pathologies, including cancer. SNO can affect the function and stability of different proteins, such as the mitochondrial chaperone TRAP1.
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