کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
5514207 1541592 2017 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Alternative fluorimetric-based method to detect and compare total S-nitrosothiols in plants
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Alternative fluorimetric-based method to detect and compare total S-nitrosothiols in plants
چکیده انگلیسی


- S-nitrosothiols can be detected by UV photolysis and fluorophore-based quantification.
- The fluorophore-based method sensitivity is about six times higher than the Saville reaction.
- The method proved to be specific, reproducible and reliable for all species tested.

Nitric oxide (NO) is an important signaling molecule occurring in virtually all organisms, whose mechanism of action relies mainly on its interaction with proteins or peptides by nitrosylation, forming compounds such as S-nitrosothiols (SNO). The Saville reaction and the ozone-based chemiluminescence method are the main techniques used for nitrosylated protein quantification. While the Saville assay is not very sensitive, the ozone-based chemiluminescence is expensive and time-consuming. Here we propose a method in which the protein-bound NO groups are exposed to UV light, cleaving the bond and allowing the quantification of the derived NO molecules by diamino-rhodamine (DAR) dyes. The DAR-based method was shown to be specific in plant tissues by pre-treatment of the samples with reducing agents and parallel EPR analysis. Spike-and-recovery assays revealed 72% recovery after a GSNO spike. Moreover, the method was significantly more sensitive than the Saville reaction, and this increase in sensitivity was crucial for detecting the reduced levels of nitrosylated proteins in plant species other than Arabidopsis. The method presented here is a suitable alternative to compare plant samples, allowing simple and fast detection of nitrosylated proteins.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Nitric Oxide - Volume 68, 1 August 2017, Pages 7-13
نویسندگان
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