TY  - JOUR
A1  - Litty, Dennis
A1  - Müller, Volker
T1  - A Na+ A1AO ATP synthase with a V-type c subunit in a mesophilic bacterium
T2  - The FEBS journal
N2  - 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.
KW  - acetogen
KW  - bioenergetics
KW  - Eubacterium
KW  - Na+ transport
Y1  - 2019
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/63825
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-638257
SN  - 1742-4658
N1  - Financial support by the Deutsche Forschungsgemeinschaft via SFB807 is gratefully acknowledged.
VL  - 287
IS  - 14
SP  - 3012
EP  - 3023
PB  - Wiley-Blackwell
CY  - Oxford [u.a.]
ER  -