A Na+ A1AO ATP synthase with a V-type c subunit in a mesophilic bacterium
- A1AO ATP synthases with a V-type c subunit have only been found in hyperthermophilic archaea which makes bioenergetic analyses impossible due to the instability of liposomes at high temperatures. A search for a potential archaeal A1AO ATP synthase with a V-type c subunit in a mesophilic organism revealed an A1AO ATP synthase cluster in the anaerobic, acetogenic bacterium Eubacterium limosum KIST612. The enzyme was purified to apparent homogeneity from cells grown on methanol to a specific activity of 1.2 U·mg−1 with a yield of 12%. The enzyme contained subunits A, B, C, D, E, F, H, a, and c. Subunit c is predicted to be a typical V-type c subunit with only one ion (Na+)-binding site. Indeed, ATP hydrolysis was strictly Na+-dependent. N,N′-dicyclohexylcarbodiimide (DCCD) inhibited ATP hydrolysis, but inhibition was relieved by addition of Na+. Na+ was shown directly to abolish binding of the fluorescence DCCD derivative, NCD-4, to subunit c, demonstrating a competition of Na+ and DCCD/NCD-4 for a common binding site. After incorporation of the A1AO ATP synthase into liposomes, ATP-dependent primary transport of 22Na+ as well as ΔµNa+-driven ATP synthesis could be demonstrated. The Na+ A1AO ATP synthase from E. limosum is the first ATP synthase with a V-type c subunit from a mesophilic organism. This will enable future bioenergetic analysis of these unique ATP synthases.
Author: | Dennis LittyGND, Volker MüllerORCiD |
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URN: | urn:nbn:de:hebis:30:3-638257 |
DOI: | https://doi.org/10.1111/febs.15193 |
ISSN: | 1742-4658 |
Parent Title (English): | The FEBS journal |
Publisher: | Wiley-Blackwell |
Place of publication: | Oxford [u.a.] |
Document Type: | Article |
Language: | English |
Date of Publication (online): | 2019/12/26 |
Date of first Publication: | 2019/12/26 |
Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
Release Date: | 2022/03/08 |
Tag: | Eubacterium; Na+ transport; acetogen; bioenergetics |
Volume: | 287 |
Issue: | 14 |
Page Number: | 12 |
First Page: | 3012 |
Last Page: | 3023 |
Note: | Financial support by the Deutsche Forschungsgemeinschaft via SFB807 is gratefully acknowledged. |
HeBIS-PPN: | 494118016 |
Institutes: | Biowissenschaften |
Dewey Decimal Classification: | 5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie |
Sammlungen: | Universitätspublikationen |
Licence (German): | Creative Commons - Namensnennung 4.0 |