Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
2020889 | Protein Expression and Purification | 2011 | 8 Pages |
Abstract
The vacuolar-type H+-ATPase (V-ATPase) is a multi-subunit proton pump that is involved in both intra- and extracellular acidification processes throughout human body. Subunits constituting the peripheral stalk of the V-ATPase are known to have several isoforms responsible for tissue/cell specific different physiological roles. To study the different interaction of these isoforms, we expressed and purified the isoforms of human V-ATPase peripheral stalk subunits using Escherichia coli cell-free protein synthesis system: E1, E2, G1, G2, G3, C1, C2, H and N-terminal soluble part of a1 and a2 isoforms. The purification conditions were different depending on the isoforms, maybe reflecting the isoform specific biochemical characteristics. The purified proteins are expected to facilitate further experiments to study about the cell specific interaction and regulation and thus provide insight into physiological meaning of the existence of several isoforms of each subunit in V-ATPase.
Keywords
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Biochemistry, Genetics and Molecular Biology
Biochemistry
Authors
Suhaila Rahman, Yoshiko Ishizuka-Katsura, Satoshi Arai, Shinya Saijo, Ichiro Yamato, Mitsutoshi Toyama, Noboru Ohsawa, Mio Inoue, Keiko Honda, Takaho Terada, Mikako Shirouzu, Shigeyuki Yokoyama, So Iwata, Takeshi Murata,