A simplified calculation procedure for mass isotopomer distribution analysis (MIDA) based on multiple linear regression
A simplified calculation procedure for mass isotopomer distribution analysis (MIDA) based on multiple linear regression
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DOI：
10.1002/jms.3809
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发表时间：
2016-10-01
影响因子：
2.3
通讯作者：
Garcia Alonso, J. Ignacio
中科院分区：
文献类型：
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作者：
Fernandez-Fernandez, Mario;Rodriguez-Gonzalez, Pablo;Garcia Alonso, J. Ignacio
我们已经开发了一种新的，快速和简单的计算程序的质量同位素分布分析的基础上，多元线性回归，允许同时计算的前体池富集和分数的新合成的标记蛋白质（分步合成）使用线性代数。为了测试这种方法，我们使用肽RGGGGLK作为包含甘氨酸的三个亚基的模型胰蛋白酶肽。我们选择标记在两个C-13原子中的甘氨酸（C-13（2）-甘氨酸）作为标记的氨基酸，以证明光谱重叠在所提出的方法中不是问题。通过将前体池富集从10%改变为40%的C-13（2）-甘氨酸，首先在体外测试所开发的方法。其次，通过将天然丰度的RGGGGLK肽和10或20%C-13（2）-甘氨酸以1：1、1：3和3：1的比例组合来设计蛋白质的模拟体内合成。使用简单的电子表格计算前体池富集和分步合成值，具有令人满意的精密度和准确度。这种新的方法可以提供一个相对快速和简单的手段来测量蛋白质周转的基础上稳定的同位素示踪剂。版权所有（c）2016约翰威利父子有限公司
We have developed a novel, rapid and easy calculation procedure for Mass Isotopomer Distribution Analysis based on multiple linear regression which allows the simultaneous calculation of the precursor pool enrichment and the fraction of newly synthesized labelled proteins (fractional synthesis) using linear algebra. To test this approach, we used the peptide RGGGLK as a model tryptic peptide containing three subunits of glycine. We selected glycine labelled in two C-13 atoms (C-13(2)-glycine) as labelled amino acid to demonstrate that spectral overlap is not a problem in the proposed methodology. The developed methodology was tested first in vitro by changing the precursor pool enrichment from 10 to 40% of C-13(2)-glycine. Secondly, a simulated in vivo synthesis of proteins was designed by combining the natural abundance RGGGLK peptide and 10 or 20% C-13(2)-glycine at 1:1, 1:3 and 3:1 ratios. Precursor pool enrichments and fractional synthesis values were calculated with satisfactory precision and accuracy using a simple spreadsheet. This novel approach can provide a relatively rapid and easy means to measure protein turnover based on stable isotope tracers. Copyright (c) 2016 John Wiley & Sons, Ltd.