RAPID, SENSITIVE ANALYSIS OF PROTEIN MIXTURES BY MASS-SPECTROMETRY
RAPID, SENSITIVE ANALYSIS OF PROTEIN MIXTURES BY MASS-SPECTROMETRY
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DOI：
10.1073/pnas.87.17.6873
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发表时间：
1990-09-01
影响因子：
11.1
通讯作者：
CHAIT, BT
中科院分区：
文献类型：
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作者：
BEAVIS, RC;CHAIT, BT
我们建立了一种用质谱仪测定复杂混合物中蛋白质分子质量的方法。该方法能够在不使用任何色谱分离步骤的情况下检测混合物的成分，并将容忍相对大量的缓冲液和无机污染物。它可以同时测定从1 kDa到40 kDa的蛋白质分子质量，准确度为.+-。0.01%和40 kDa以上，准确度降低。实际检测蛋白质的下限是0.1µm的浓度，并且消耗1µl的这种溶液。分析速度非常快：<；15分钟即可完成完整的分析，包括样品准备、质谱计导入、质谱图采集和数据整理。获得的质谱图不需要详细的解释，因为没有电离蛋白质(或蛋白质亚单位)分子的碎裂。因此，在质谱峰和原始混合物中存在的蛋白质之间存在一一对应关系。光谱假定为色谱图的外观，横坐标是质量电荷比而不是色层保留时间。
We have developed a method for determining the molecular masses of proteins in complex mixtures by mass spectrometry. The method has the capacity to examine the components of mixtures without using any chromatographic separation steps and will tolerate relatively large amounts of buffers and inorganic contaminants. It allows the simultaneous determination of protein molecular masses from 1 to 40 kDa with an accuracy of .+-. 0.01% and above 40 kDa with reduced accuracy. The lower limit for practical detection of a protein is a concentration of .apprxeq.0.1 .mu.M, and < 1 .mu.l of such a solution is consumed. The analysis is very fast: < 15 min is necessary to perform the complete analysis, including sample preparation, introduction into the mass spectrometer, mass spectrum collection, and data reduction. The mass spectrum that is obtained does not require elaborate interpretation because there is no fragmentation of the ionized protein (or protein subunit) molecule. Therefore, there is a one-to-one correspondence between the peaks in the mass spectrum and the proteins present in the original mixture. The spectra assume the appearance of chromatograms, with the abscissa being mass-to-charge ratio rather than chromatographic retention time.