title: Binding interactions of the peripheral stalk subunit isoforms from human V-ATPase.
abstract: The mammalian peripheral stalk subunits of the vacuolar-type H(+)-ATPases (V-ATPases) possess several isoforms (C1, C2, E1, E2, G1, G2, G3, a1, a2, a3, and a4), which may play significant role in regulating ATPase assembly and disassembly in different tissues. To better understand the structure and function of V-ATPase, we expressed and purified several isoforms of the human V-ATPase peripheral stalk: E1G1, E1G2, E1G3, E2G1, E2G2, E2G3, C1, C2, H, a1NT, and a2NT. Here, we investigated and characterized the isoforms of the peripheral stalk region of human V-ATPase with respect to their affinity and kinetics in different combination. We found that different isoforms interacted in a similar manner with the isoforms of other subunits. The differences in binding affinities among isoforms were minor from our in vitro studies. However, such minor differences from the binding interaction among isoforms might provide valuable information for the future structural-functional studies of this holoenzyme.
authors: a Department of Biological Science and Technology , Tokyo University of Science , Tokyo , Japan.
authors: Suhaila
authors: S
authors: Rahman
authors: a Department of Biological Science and Technology , Tokyo University of Science , Tokyo , Japan.
authors: Ichiro
authors: I
authors: Yamato
authors: a Department of Biological Science and Technology , Tokyo University of Science , Tokyo , Japan.
authors: Shinya
authors: S
authors: Saijo
authors: a Department of Biological Science and Technology , Tokyo University of Science , Tokyo , Japan.|b Department of Chemistry , Graduate School of Science, Chiba University , Chiba , Japan.
authors: Kenji
authors: K
authors: Mizutani
authors: b Department of Chemistry , Graduate School of Science, Chiba University , Chiba , Japan.
authors: Yuuki
authors: Y
authors: Takamuku
authors: c RIKEN Systems and Structural Biology Center , Yokohama , Japan.
authors: Yoshiko
authors: Y
authors: Ishizuka-Katsura
authors: c RIKEN Systems and Structural Biology Center , Yokohama , Japan.
authors: Noboru
authors: N
authors: Ohsawa
authors: c RIKEN Systems and Structural Biology Center , Yokohama , Japan.
authors: Takaho
authors: T
authors: Terada
authors: c RIKEN Systems and Structural Biology Center , Yokohama , Japan.
authors: Mikako
authors: M
authors: Shirouzu
authors: c RIKEN Systems and Structural Biology Center , Yokohama , Japan.|d Department of Biophysics and Biochemistry , Graduate School of Science, The University of Tokyo , Tokyo , Japan.
authors: Shigeyuki
authors: S
authors: Yokoyama
authors: b Department of Chemistry , Graduate School of Science, Chiba University , Chiba , Japan.|c RIKEN Systems and Structural Biology Center , Yokohama , Japan.|e Molecular Chirality Research Center, Chiba University , Chiba , Japan.|f JST, PRESTO , Chiba , Japan.
authors: Takeshi
authors: T
authors: Murata
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002474:Cell-Free System / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D011485:Protein Binding; D014176:Protein Biosynthesis; D054730:Protein Interaction Domains and Motifs; D055503:Protein Multimerization; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D025262:Vacuolar Proton-Translocating ATPases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D007527:Isoenzymes; D021122:Protein Subunits; D011994:Recombinant Proteins; D025262:Vacuolar Proton-Translocating ATPases
keywords: V-ATPase; affinity; human peripheral stalk; subunit isoform; surface plasmon resonance