Bacterial F-type ATP synthases follow a well-choreographed assembly pathway.

Vu Huu, Khanh; Zangl, Rene; Hoffmann, Jan; Just, Alicia; Morgner, Nina · Nat Commun · 2022

basic_science · Level V

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Abstract

F-type ATP synthases are multiprotein complexes composed of two separate coupled motors (F<sub>1</sub> and F<sub>O</sub>) generating adenosine triphosphate (ATP) as the universal major energy source in a variety of relevant biological processes in mitochondria, bacteria and chloroplasts. While the structure of many ATPases is solved today, the precise assembly pathway of F<sub>1</sub>F<sub>O</sub>-ATP synthases is still largely unclear. Here, we probe the assembly of the F<sub>1</sub> complex from Acetobacterium woodii. Using laser induced liquid bead ion desorption (LILBID) mass spectrometry, we study the self-assembly of purified F<sub>1</sub> subunits in different environments under non-denaturing conditions. We report assembly requirements and identify important assembly intermediates in vitro and in cellula. Our data provide evidence that nucleotide binding is crucial for in vitro F<sub>1</sub> assembly, whereas ATP hydrolysis appears to be less critical. We correlate our results with activity measurements and propose a model for the assembly pathway of a functional F<sub>1</sub> complex.

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