Essential arginine in subunit a and aspartate in subunit c of FoF1 ATP synthase: effect of repositioning within helix 4 of subunit a and helix 2 of subunit c.
FoF1 ATP synthase couples proton flow through the integral membrane portion Fo (ab2c10) to ATP-synthesis in the extrinsic F1-part ((alphabeta)3gammadeltaepsilon) (Escherichia coli nomenclature and stoichiometry). Coupling occurs by mechanical rotation of subunits c10gammaepsilon relative to (alphabeta)3deltaab2. Two residues were found to be essential for proton flow through ab2c10, namely Arg210 in subunit a (aR210) and Asp61 in subunits c (cD61). Their deletion abolishes proton flow, but "horizontal" repositioning, by anchoring them in adjacent transmembrane helices, restores function. Here, we investigated the effects of "vertical" repositioning aR210, cD61, or both by one helical turn towards the N- or C-termini of their original helices. Other than in the horizontal the vertical displacement changes the positions of the side chains within the depth of the membrane. Mutant aR210A/aN214R appeared to be short-circuited in that it supported proton conduction only through EF1-depleted EFo, but not in EFoEF1, nor ATP-driven proton pumping. Mutant cD61N/cM65D grew on succinate, retained the ability to synthesize ATP and supported passive proton conduction but apparently not ATP hydrolysis-driven proton pumping.
Langemeyer
Lars
L
Universität Osnabrück, Fachbereich Biologie, Biochemie, Barbarastr. 13, 49076 Osnabrück, Germany.
Engelbrecht
Siegfried
S
D000255:Adenosine Triphosphate / Q000378:metabolism; D001120:Arginine / Q000737:chemistry* / Q000235:genetics; D001224:Aspartic Acid / Q000737:chemistry* / Q000235:genetics; D025243:Bacterial Proton-Translocating ATPases / Q000737:chemistry* / Q000235:genetics; D002462:Cell Membrane / Q000201:enzymology; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D006868:Hydrolysis; D017433:Protein Structure, Secondary; D017494:Proton Pumps / Q000737:chemistry* / Q000235:genetics; D019802:Succinic Acid / Q000378:metabolism
D017494:Proton Pumps; D001224:Aspartic Acid; D000255:Adenosine Triphosphate; D001120:Arginine; D019802:Succinic Acid; D025243:Bacterial Proton-Translocating ATPases