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无细胞蛋白质合成反应中用于ATP再生的能量系统。

Energy systems for ATP regeneration in cell-free protein synthesis reactions.

作者信息

Calhoun Kara A, Swartz James R

机构信息

Department of Chemical Engineering, Stanford University, Stanford, CA, USA.

出版信息

Methods Mol Biol. 2007;375:3-17. doi: 10.1007/978-1-59745-388-2_1.

Abstract

Supplying energy for cell-free protein synthesis reactions is one of the biggest challenges to the success of these systems. Oftentimes, short reaction duration is attributed to an unstable energy source. Traditional cell-free reactions use a compound with a high-energy phosphate bond, such as phosphoenolpyruvate, to generate the ATP required to drive transcription and translation. However, recent work has led to better understanding and activation of the complex metabolism that can occur during cell-free reactions. We are now able to generate ATP using energy sources that are less expensive and more stable. These energy sources generally involve multistep enzymatic reactions or recreate entire energy-generating pathways, such as glycolysis and oxidative phosphorylation. We describe the various types of energy sources used in cell-free reactions, give examples of the major classes, and demonstrate protocols for successful use of three recently developed energy systems: PANOxSP, cytomim, and glucose.

摘要

为无细胞蛋白质合成反应提供能量是这些系统成功面临的最大挑战之一。通常,反应持续时间短归因于能量来源不稳定。传统的无细胞反应使用具有高能磷酸键的化合物,如磷酸烯醇丙酮酸,来生成驱动转录和翻译所需的ATP。然而,最近的研究使人们对无细胞反应过程中可能发生的复杂代谢有了更好的理解和激活。我们现在能够使用成本更低且更稳定的能量来源来生成ATP。这些能量来源通常涉及多步酶促反应或重建整个能量生成途径,如糖酵解和氧化磷酸化。我们描述了无细胞反应中使用的各种能量来源,给出了主要类别示例,并展示了成功使用三种最近开发的能量系统(PANOxSP、细胞模拟物和葡萄糖)的方案。

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