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PMID:1311852
| Citation |
Sarafian, V, Kim, Y, Poole, RJ and Rea, PA (1992) Molecular cloning and sequence of cDNA encoding the pyrophosphate-energized vacuolar membrane proton pump of Arabidopsis thaliana. Proc. Natl. Acad. Sci. U.S.A. 89:1775-9 |
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| Abstract |
The energy-dependent transport of solutes across the vacuolar membrane (tonoplast) of plant cells is driven by two H+ pumps: a vacuolar ("V-type") H(+)-ATPase (EC 3.6.1.3) and a H(+)-translocating (pyrophosphate-energized) inorganic pyrophosphatase (H(+)-PPase; EC 3.6.1.1). The H(+)-PPase, like the V-type H(+)-ATPase, is abundant and ubiquitous in the vacuolar membranes of plant cells, and both enzymes make a substantial contribution to the transtonoplast H(+)-electrochemical potential difference. Here, we report the cloning and sequence of cDNAs encoding the tonoplast H(+)-PPase of Arabidopsis thaliana. The protein predicted from the nucleotide sequence of the cDNAs is constituted of 770 amino acids and has a molecular weight of 80,800. It is a highly hydrophobic integral membrane protein, and the structure derived from hydrophilicity plots contains at least 13 transmembrane spans. Since the tonoplast H(+)-PPase appears to be constituted of one polypeptide species and genomic Southern analyses indicate that the gene encoding the Mr 80,800 polypeptide is present in only a single copy in the genome of Arabidopsis, it is suggested that the H(+)-PPase has been cloned in its entirety. The lack of sequence identities between the tonoplast H(+)-PPase and any other characterized H+ pump or PPi-dependent enzyme implies a different evolutionary origin for this translocase. |
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| Keywords |
Amino Acid Sequence; Base Sequence; Biological Transport, Active; Cloning, Molecular; DNA/genetics; Diphosphates/metabolism; Gene Expression; Genes, Plant; Hydrogen-Ion Concentration; Inorganic Pyrophosphatase; Intracellular Membranes/metabolism; Membrane Proteins/genetics; Molecular Sequence Data; Plants/genetics; Pyrophosphatases/genetics; RNA, Messenger/genetics; Restriction Mapping; Solubility; Vacuoles/metabolism; Vacuoles/ultrastructure |
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Significance
Annotations
| Gene product | Qualifier | GO ID | GO term name | Evidence Code | with/from | Aspect | Notes | Status |
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