The inner workings of an enzyme
The inner workings of an enzyme
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酶的内部工作原理
DOI：
10.1126/science.abj8346
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发表时间：
2021
期刊：
影响因子：
56.9
通讯作者：
Whitehead, Timothy A.
中科院分区：
文献类型：
--
作者：
Baumer, Zachary T.;Whitehead, Timothy A.
预测了解特定氨基酸序列如何编码酶功能是分子生物学中的一项重大挑战，对从工业生物技术，计算蛋白质设计和农业到疾病突变的预测鉴定（1）和药物化学等领域产生深远影响。为了实现这一目标，需要对酶功能的不同方面进行高通量和定量测量的创新方法。第411页，Markinet等人。(2)描述了一个名为高通量微流体酶动力学（HT-MEK）的芯片实验室平台，作为朝着这个方向迈出的一步。该技术允许高保真度的1000多个突变体的模型酶PafA（磷酸盐抑制碱性磷酸酶的黄杆菌）的体外生化和生物物理特性。HT-MEK识别了部分重叠但不同的氨基酸网络，这些网络构成了PafA的各个反应步骤，阐明了这种酶催化的机理基础。
Predictive understanding for how a particular amino acid sequence encodes enzymatic function is a grand challenge in molecular biology, with profound impacts in fields ranging from industrial biotechnology, computational protein design, and agriculture to predictive identification of disease mutations (1) and medicinal chemistry. Innovative methods for high-throughput and quantitative measurements of different aspects of enzymatic function are needed to achieve this goal. On page 411 of this issue, Markinet al.(2) describe a laboratory-on-a-chip platform called High-Throughput Microfluidic Enzyme Kinetics (HT-MEK) as a step in this direction. The technique allows high-fidelity in vitro biochemical and biophysical characterization of more than 1000 mutants of the model enzyme PafA (phosphate-irrepressible alkaline phosphatase of Flavobacterium). HT-MEK identifies partially overlapping yet distinct networks of amino acids that undergird individual reaction steps of PafA, illuminating the mechanistic basis of catalysis for this enzyme.
DOI：
10.1126/science.abf8761
发表时间：
2021-07-23
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Markin CJ;Mokhtari DA;Sunden F;Appel MJ;Akiva E;Longwell SA;Sabatti C;Herschlag D;Fordyce PM
通讯作者：
Fordyce PM