Fang Ji-Kang, Prabu Subbuswamy K, Sepuri Naresh B, Raza Haider, Anandatheerthavarada Hindupur K, Galati Domenico, Spear Joseph, Avadhani Narayan G
Laboratory of Biochemistry, The Department of Animal Biology, School of Veterinary Medicine, 3800 Spruce Street, University of Pennsylvania, Philadelphia, PA 19104, United States.
FEBS Lett. 2007 Apr 3;581(7):1302-10. doi: 10.1016/j.febslet.2007.02.042. Epub 2007 Feb 28.
We have mapped the sites of ischemia/reperfusion-induced phosphorylation of cytochrome c oxidase (CcO) subunits in rabbit hearts by using a combination of Blue Native gel/Tricine gel electrophoresis and nano-LC-MS/MS approaches. We used precursor ion scanning combined with neutral loss scanning and found that mature CcO subunit I was phosphorylated at tandem Ser115/Ser116 positions, subunit IVi1 at Thr52 and subunit Vb at Ser40. These sites are highly conserved in mammalian species. Molecular modeling suggests that phosphorylation sites of subunit I face the inter membrane space while those of subunits IVi1 and Vb face the matrix side.
我们通过结合蓝色天然凝胶/Tris-甘氨酸凝胶电泳和纳升液相色谱-串联质谱法，绘制了兔心脏中缺血/再灌注诱导的细胞色素c氧化酶（CcO）亚基磷酸化位点图谱。我们使用前体离子扫描结合中性丢失扫描，发现成熟的CcO亚基I在Ser115/Ser116串联位点磷酸化，亚基IVi1在Thr52磷酸化，亚基Vb在Ser40磷酸化。这些位点在哺乳动物物种中高度保守。分子建模表明，亚基I的磷酸化位点面向膜间隙，而亚基IVi1和Vb的磷酸化位点面向基质侧。
