High-Resolution HDX-MS of Cytochrome c Using Pepsin/Fungal Protease Type XIII Mixed Bed Column
High-Resolution HDX-MS of Cytochrome c Using Pepsin/Fungal Protease Type XIII Mixed Bed Column
复制标题
使用胃蛋白酶/真菌蛋白酶 XIII 型混合床柱对细胞色素 c 进行高分辨率 HDX-MS
DOI：
10.1007/s13361-018-2087-7
复制
发表时间：
2018
影响因子：
3.2
通讯作者：
Terry Zhang
中科院分区：
文献类型：
--
作者：
Y. Hamuro;Terry Zhang
摘要：与单纯的pepsin色谱柱相比，pepsin/FPXIII（来自Aspergillus saiitoi的蛋白酶，XIII型）混合床色谱柱可显著提高自下而上氢/氘交换质谱（HDX-MS）数据的分辨率。采用混合床柱的HDX-MS法测定了100个细胞色素c主酰胺氢中65个酰胺氢的交换率。两种酶的不同裂解特性产生了138个独特的高质量的肽片段，从而实现了氘的精细亚定位。在这种方法中确定的汇率是一致的，在当前的研究以及与先前的HDX-NMR研究。高分辨率HDX-MS数据可以确定每个残留物的交换率，而不是消化片段的氘积累曲线。汇率以更可复制的方式提供了更精确和定量的蛋白质动力学测量。
AbstractA pepsin/FPXIII (protease from Aspergillus saitoi, type XIII) mixed bed column significantly improved the resolution of bottom-up hydrogen/deuterium exchange mass spectrometry (HDX-MS) data compared with a pepsin-only column. The HDX-MS method using the mixed bed column determined 65 amide hydrogen exchange rates out of one hundred cytochrome c backbone amide hydrogens. Different cleavage specificities of the two enzymes generated 138 unique high-quality peptic fragments, which allows fine sub-localization of deuterium. The exchange rates determined in this method are consistent within the current study as well as with the previous HDX-NMR study. High-resolution HDX-MS data can determine the exchange rate of each residue not the deuterium buildup curve of a peptic fragment. The exchange rates provide more precise and quantitative measurements of protein dynamics in a more reproducible manner.
Graphical Abstractᅟ
影响因子：
7.4
作者：
Zhang, Hui-Min;Kazazic, Sasa;Schaub, Tanner M.;Tipton, Jeremiah D.;Emmett, Mark R.;Marshall, Alan G.
通讯作者：
Marshall, Alan G.
影响因子：
7.4
作者：
Sterling, Harry J.;Williams, Evan R.
通讯作者：
Williams, Evan R.
影响因子：
2.9
作者：
Huang, Richard Y-C.;Garai, Kanchan;Frieden, Carl;Gross, Michael L.
通讯作者：
Gross, Michael L.