Summary
S-nitrosylation, or S-nitrosation, is an important post-translational modification of proteins. It is conducted by the connection of NO, its derivative and cysteine thiol groups on the protein. It is also the most important channel for biological effects of nitric oxide signaling molecule. More and more researches show that S-nitrosylation plays a very significant role in cell signal transduction and diseases diagnosis.
The main factor affecting the biological function of the modified is the lack of proper s-nitrosylation analysis of protein. Discovering unknown nitrosylation protein, determining nitrosylation sites, getting the quantitative change information of nitrosylation and the dynamic changes in biological processes are the main projects, which are meaningful for the biological effects and molecular mechanisms of nitrosylation. In this article, we are going to study the achievements gained in this process.
1.Using the specific fluorescent probe---AMCA-HPDP to bind with SDS-PAGE, through which a method for detecting the identification of single exogenous protein nitrosylation is formed. This method is called AMCA-Switch. Compared with the widely used Biotin-Switch method, fluorescence with in-gel detecting method is more intuitive than the Western Blot, saving 3 or 4 experimental hours and avoiding the interference of endogenous biotinylated proteins. What’s more, as for the problem of biotin-avidin purification system, the thiol-containing peptides with organic fluorine probe can be designed and synthesized with low cost, and good specificity. This offers a great purification method for high-throughput genomics.
2.Based on the Biotin-Switch method, S-nitrosylation uses the Urea as the mercapto closed and labeled environment and replaces the protein purification with peptide segment purification. Combined with LC-MS/MS technology, the high-throutput method to detect targets and sites of the modified protein nitrosylation. Compared with the existed method---SNOSID, this method releases the detergents’ inhibition against ESI-LC-MS/MS signal, which repeats the result. Setting proper comparison and describing the changes of nitroso modification content in each sample group, this method avoids the false-positive results caused by Vc.
These are two main achievements for the s-nitrosylation analysis of protein. In Creative Proteomics, we can offer high-quality s-nitrosylation analysis of protein for our customers.
S-nitrosylation, or S-nitrosation, is an important post-translational modification of proteins. It is conducted by the connection of NO, its derivative and cysteine thiol groups on the protein. It is also the most important channel for biological effects of nitric oxide signaling molecule. More and more researches show that S-nitrosylation plays a very significant role in cell signal transduction and diseases diagnosis.
The main factor affecting the biological function of the modified is the lack of proper s-nitrosylation analysis of protein. Discovering unknown nitrosylation protein, determining nitrosylation sites, getting the quantitative change information of nitrosylation and the dynamic changes in biological processes are the main projects, which are meaningful for the biological effects and molecular mechanisms of nitrosylation. In this article, we are going to study the achievements gained in this process.
1.Using the specific fluorescent probe---AMCA-HPDP to bind with SDS-PAGE, through which a method for detecting the identification of single exogenous protein nitrosylation is formed. This method is called AMCA-Switch. Compared with the widely used Biotin-Switch method, fluorescence with in-gel detecting method is more intuitive than the Western Blot, saving 3 or 4 experimental hours and avoiding the interference of endogenous biotinylated proteins. What’s more, as for the problem of biotin-avidin purification system, the thiol-containing peptides with organic fluorine probe can be designed and synthesized with low cost, and good specificity. This offers a great purification method for high-throughput genomics.
2.Based on the Biotin-Switch method, S-nitrosylation uses the Urea as the mercapto closed and labeled environment and replaces the protein purification with peptide segment purification. Combined with LC-MS/MS technology, the high-throutput method to detect targets and sites of the modified protein nitrosylation. Compared with the existed method---SNOSID, this method releases the detergents’ inhibition against ESI-LC-MS/MS signal, which repeats the result. Setting proper comparison and describing the changes of nitroso modification content in each sample group, this method avoids the false-positive results caused by Vc.
These are two main achievements for the s-nitrosylation analysis of protein. In Creative Proteomics, we can offer high-quality s-nitrosylation analysis of protein for our customers.