Study on acidic environment formation by V-ATPase using single molecule observation

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We have found that osteoclast-specific V-ATPase(vacuole-type proton-pumping ATPase) is composed of a3 and d2 isoforms. V-ATPase with a3 isoform is localized on lysosomal membrane, and is essential for traffic of lysosomes to plasmamembrane. Secretion of lysosomal enzymes and acidification outside osteoclasts are important for bone resorption. On the other hand, we have established experimental systems to observe subunit rotation of single molecule V-ATPase and to examine assembly of the catalytic domain and the proton pathway. These systems are essential to elucidate molecular mechanism that creates various acidic environments.