NO引起Fyn酪氨酸激酶巯基亚硝基化的体外研究

Nitric oxide causes the S - nitrosylation of Fyn tyrosine kinase in vitro

目的 探讨离体情况下一氧化氮（NO）对酪氨酸蛋白激酶Fyn蛋白巯基亚硝基化的影响.方法 HEK293细胞转染pcDNA3.1-Fyn质粒后,用S-亚硝基谷胱甘肽（GSNO）刺激,或共转染神经型一氧化氮合酶（nNOS）-Fyn质粒,再用A23187和7-硝基吲唑（7-NI）刺激,生物素转换法（biotin-switch method）和Western blotting检测Fyn巯基亚硝基化情况.结果 GSNO以及 nNOS在A23187存在的情况下,都能够引起Fyn巯基亚硝基化.结论外源性和内源性NO都能够使Fyn巯基亚硝基化.

Objective To investigate the effect of nitric oxide （NO） on the S - nitrosylation of Fyn kinase in vitro. Methods HEK293 cell was transfected with pcDNA3.1 - Fyn plasmid and then was stimulated with S - nitrosoglutathione （GSNO）, or co -transfected with inducible nitric oxide synthase （nNOS） -Fyn plasmid and then was stimulated with A23187 and 7 - nitroindazole （7 - NI）, the S - nitrosylation of Fyn was examined by biotin - switch method and Western blotting. Results Both GSNO and nNOS with A23187 could cause the S -nitrosylation of Fyn. Conclusion Both ectogenic and endogenic NO can cause the S - nitrosylation of Fyn.