Physical interaction between aldolase and vacuolar H+-ATPase is essential for the assembly and activity of the proton pump.
Vacuolar proton-translocating ATPases (V-ATPases) are a family of highly conserved proton pumps that couple hydrolysis of cytosolic ATP to proton transport out of the cytosol. Although V-ATPases are involved in a number of cellular processes, how the proton pumps are regulated under physiological conditions is not well understood. We have reported that the glycolytic enzyme aldolase mediates V-ATPase assembly and activity by physical association with the proton pump (Lu, M., Holliday, L. S., Zhang, L., Dunn, W. A., and Gluck, S. L. (2001) J. Biol. Chem. 276, 30407-30413 and Lu, M., Sautin, Y., Holliday, L. S., and Gluck, S. L. (2004) J. Biol. Chem. 279, 8732-8739). In this study, we generate aldolase mutants that lack binding to the B subunit of V-ATPase but retain normal catalytic activities. Functional analysis of the aldolase mutants shows that disruption of binding between aldolase and the B subunit of V-ATPase results in disassembly and malfunction of V-ATPase. In contrast, aldolase enzymatic activity is not required for V-ATPase assembly. Taken together, these findings strongly suggest an important role for physical association between aldolase and V-ATPase in the regulation of the proton pump.
Lu
Ming
M
Department of Medicine, University of California School of Medicine, San Francisco, California 94143-0532, USA. minglu@medicine.ucsf.edu
Ammar
David
D
Ives
Harlan
H
Albrecht
Fred
F
Gluck
Stephen L
SL
D002384:Catalysis; D005634:Fructose-Bisphosphate Aldolase / Q000737:chemistry / Q000378:metabolism*; D005656:Fungal Proteins / Q000737:chemistry; D006801:Humans; D006868:Hydrolysis; D008954:Models, Biological; D016296:Mutagenesis; D009154:Mutation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D017494:Proton Pumps / Q000378:metabolism*; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000378:metabolism*; D011522:Protons; D012441:Saccharomyces cerevisiae / Q000378:metabolism
D005656:Fungal Proteins; D017494:Proton Pumps; D011522:Protons; D006180:Proton-Translocating ATPases; D005634:Fructose-Bisphosphate Aldolase