학술논문

Isolation of intact and active FoF1 ATP synthase using a FLAG-tagged subunit from the cyanobacterium Synechocystis sp. PCC 6803
Document Type
article
Source
STAR Protocols, Vol 3, Iss 3, Pp 101623- (2022)
Subject
Metabolism
Microbiology
Plant sciences
Molecular Biology
Protein Biochemistry
Protein expression and purification
Science (General)
Q1-390
Language
English
ISSN
2666-1667
Abstract
Summary: 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 Synechocystis 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). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.