Fluorimetric-Based Method to Detect and Quantify Total S-Nitrosothiols (SNOs) in Plant Samples

Methods Mol Biol. 2020:2057:37-43. doi: 10.1007/978-1-4939-9790-9_4.

Abstract

Accumulating experimental evidence indicates that S-nitrosylation (technically S-nitrosation) events have a central role in plant biology, presumably accounting for much of the widespread influence of nitric oxide (NO) on developmental, metabolic, and stress-related plant responses. Therefore, the accurate detection and quantification of S-nitrosylated proteins and peptides can be particularly useful to determine the relevance of this class of compounds in the ever-increasing number of NO-dependent signaling events described in plant systems. Up to now, the quantification of S-nitrosothiols (SNOs) in plant samples has mostly relied on the Saville reaction and the ozone-based chemiluminescence method, which lacks sensitivity and are very time-consuming, respectively. Taking advantage of the photolytic properties of S-nitrosylated proteins and peptides, the method described in this chapter allows simple, fast, and high-throughput detection of SNOs in plant samples.

Keywords: Nitric oxide; Nitrosylated proteins; Ozone chemiluminescence; S-nitrosoglutathione; S-nitrosothiols; Saville.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fluorometry / instrumentation
  • Fluorometry / methods*
  • Luminescent Measurements / methods
  • Nitric Oxide / metabolism*
  • Nitric Oxide / radiation effects
  • Nitrites / chemistry
  • Nitrosation
  • Plant Proteins / metabolism*
  • Plants / chemistry
  • Plants / metabolism*
  • Rhodamines / chemistry
  • Rhodamines / radiation effects
  • S-Nitrosoglutathione / metabolism
  • S-Nitrosothiols / analysis*
  • Ultraviolet Rays
  • Workflow

Substances

  • Nitrites
  • Plant Proteins
  • Rhodamines
  • S-Nitrosothiols
  • diaminorhodamine-4M
  • Nitric Oxide
  • S-Nitrosoglutathione