Studies of protein-protein association between yeast cytochrome c peroxidase and yeast iso-1 ferricytochrome c by hydrogen-deuterium exchange labeling and proton NMR spectroscopy.
Studies of protein-protein association between yeast cytochrome c peroxidase and yeast iso-1 ferricytochrome c by hydrogen-deuterium exchange labeling and proton NMR spectroscopy.
复制标题
通过氢-氘交换标记和质子核磁共振波谱研究酵母细胞色素 c 过氧化物酶和酵母 iso-1 铁细胞色素 c 之间的蛋白质-蛋白质关联。
DOI：
10.1021/bi00206a004
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发表时间：
1994
期刊：
影响因子：
2.9
通讯作者：
Satterlee,JD
中科院分区：
文献类型：
--
作者：
Yi,Q;Erman,JE;Satterlee,JD
Revised Manuscript Received June 28, 19949 abstract: Hydrogen-deuterium (HD) exchange labeling and proton NMR have been applied to study the protein-protein association between cytochrome c peroxidase (CcP) and yeast iso-1 ferricytochrome c. Specifically, theexchange behavior of individual backbone amide protons of yeast iso-1 ferricytochrome c in both CcP-bound (ie, complexed) and free (ie, never in the complex) forms has been investigated and used in an attempt to map the binding site of CcP on yeast iso-1 ferricytochrome c when the noncovalent complex was formed in very low salt solution. The exchange rates of certainamino acid amide protons were significantly slowed down, by up to 40-fold, in the complex comparedto the free form. The protected regions on iso-1 ferricytochrome c include parts of the 10’s helix and the 70’s helix surrounding the cytochrome c heme solvent-exposed edge (the so-called “front side” of iso-1 cytochrome c). These regions are very similar to the cytochrome c peroxidase binding interface on iso-1 ferricytochrome c that has been defined by X-ray crystallographic data. Thisfurther supports the direct involvement of the front side of iso-1 cytochrome c in binding with cytochrome c peroxidase. The results from our HD exchange experiments also indicated that the amide proton exchange rates of Trp59, Asp60, and part of the 90’s helix, all of which are located on the opposite side (the “back” side) of ferricytochrome c from theheme solvent-exposed edge, are also retarded upon complex formation.
