Screening of combinatorial libraries for substrate preference by mass spectrometry.
Screening of combinatorial libraries for substrate preference by mass spectrometry.
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通过质谱法筛选组合文库的底物偏好。
DOI：
10.1021/ac0489925
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发表时间：
2005
影响因子：
7.4
通讯作者：
Prokai,Laszlo
中科院分区：
文献类型：
--
作者：
StevensJr,StanleyM;Prokai-Tatrai,Katalin;Prokai,Laszlo
我们提出了一种结合化学和质谱学的快速检测酶专一性的方法，并以此为例，测定了肽甘氨酸α-酰胺化酶的底物专一性，并与传统的定量技术进行了比较。尽管文库筛选的替代方法通常仅限于某些酶，并且在潜在底物的合成或衍生化方面存在困难，但我们称为基于手性的底物(CHILL)文库的同位素标记方法并不能满足这些限制，因为我们利用酶固有的立体特异性来确定首选底物。此外，与需要繁琐的方法开发的典型筛选程序相比，Chills方法可以产生准确的结果，因为合成的文库包含每个单独文库组分的结构相似的内部标准，以便定量酶反应的进展。
We present a rapid screening method for monitoring enzyme specificity using both combinatorial chemistry and mass spectrometry where, as an example, the substrate specificity of peptidylglycine α-amidating enzyme was determined and compared against a conventional quantitative technique. Whereas alternative methods for library screening are generally limited to certain enzymes and can present difficulties in the synthesis or derivatization of potential substrates, the approach we call chirality-based isotope labeling for a library of substrates (CHILLS) does not fall short to such limitations, since we exploit the inherent stereospecificity of enzymes to determine preferred substrates. Additionally, the CHILLS method generates accurate results, as compared to typical screening procedures that require tedious method development, because the synthesized library contains a structurally similar internal standard for each individual library component in order to quantitate the progress of enzymatic reactions.