使用来自蓝藻 sp. PCC 6803 的带有 FLAG 标签的亚基分离完整且活性的 FoF1 ATP 合酶。

Genetics and Experimental Bioinformatics, Faculty of Biology, University of Freiburg, D-79104 Freiburg, Germany.

Institute for Surgical Pathology, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany; Proteomic Core Facility (ProtCF), Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany.

STAR Protoc. 2022 Aug 16;3(3):101623. doi: 10.1016/j.xpro.2022.101623. eCollection 2022 Sep 16.

The FoF1 ATP synthase (ATPase) is one of the most important protein complexes in energy metabolism. The isolation of functional ATPase complexes is fundamental to address questions about its assembly, regulation, and functions. This protocol describes the purification of intact and active ATPase from the model cyanobacterium sp. PCC 6803. Basis for purification is a 3×FLAG tag fused to the beta subunit. The ATPase is enzymatically active and its purity is demonstrated using mass spectrometry, denaturing, and blue-native PAGE. For complete details on the use and execution of this protocol, please refer to Song et al. (2022).

FoF1 ATP 合酶（ATPase）是能量代谢中最重要的蛋白质复合物之一。分离具有功能的 ATPase 复合物对于解决其组装、调节和功能问题至关重要。本方案描述了从模式蓝藻 sp. PCC 6803 中纯化完整且有活性的 ATPase 的方法。纯化的基础是融合到β亚基上的 3×FLAG 标签。ATPase 具有酶活性，其纯度可通过质谱、变性和蓝色 native PAGE 进行证明。有关该方案使用和执行的完整详细信息，请参阅 Song 等人。（2022 年）。