@article{2e2563c47b194d81b050a845b9893ccd,
title = "Overexpression, purification, enzymatic and microscopic characterization of recombinant mycobacterial F-ATP synthase",
abstract = "The F-ATP synthase is an essential enzyme in mycobacteria, including the pathogenic Mycobacterium tuberculosis. Several new compounds in the TB-drug pipeline target the F-ATP synthase. In light of the importance and pharmacological attractiveness of this novel antibiotic target, tools have to be developed to generate a recombinant mycobacterial F1FO ATP synthase to achieve atomic insight and mutants for mechanistic and regulatory understanding as well as structure-based drug design. Here, we report the first genetically engineered, purified and enzymatically active recombinant M. smegmatis F1FO ATP synthase. The projected 2D- and 3D structures of the recombinant enzyme derived from negatively stained electron micrographs are presented. Furthermore, the first 2D projections from cryo-electron images are revealed, paving the way for an atomic resolution structure determination.",
keywords = "Bioenergetics, Drug discovery, Electron microscopy, F-ATP synthase, Mycobacterium, Tuberculosis",
author = "Saw, {Wuan Geok} and Wong, {Chui Fann} and Thomas Dick and Gerhard Gr{\"u}ber",
note = "Funding Information: The authors acknowledge the use of the EM facility at the NTU Institute of Structural Biology. The research and the PhD scholarship of C.-F. Wong were supported by the National Research Foundation ( NRF ) Singapore, NRF Competitive Research Programme (CRP), Grant Award Number NRF–CRP18–2017–01 . Funding Information: In October 2019, a $14 Bi Global Fund to fight Tuberculosis (TB), Malaria and Aids was initiated [1] and the WHO reported 1.4 million deaths in the 2018 due to TB, reflecting the need to tackle these important disease [2]. The current treatment of TB mainly utilizes a combination of antibiotics that requires long-term therapy [3]. An additional problem to treat TB lies in the emergence of multidrug-resistant TB (MDR-TB) and extensively drug-resistant TB (XDR-TB) [4,5].The authors acknowledge the use of the EM facility at the NTU Institute of Structural Biology. The research and the PhD scholarship of C.-F. Wong were supported by the National Research Foundation (NRF) Singapore, NRF Competitive Research Programme (CRP), Grant Award Number NRF?CRP18?2017?01. Publisher Copyright: {\textcopyright} 2019 Elsevier Inc.",
year = "2020",
month = feb,
day = "5",
doi = "10.1016/j.bbrc.2019.11.098",
language = "English (US)",
volume = "522",
pages = "374--380",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "2",
}