Tissue specificity of E subunit isoforms of plant vacuolar H(+)-ATPase and existence of isotype enzymes.
Immunoblot analyses and partial amino acid sequencings revealed that both the 40- (E1) and 37-kDa (E2) subunits of V-ATPase in the pea epicotyl were E subunit isoforms. Similarly, both the 35- (D1) and 29-kDa (D2) subunits were D subunit isoforms, although the similarity of the amino acid sequences is still unknown. In immunoblot analyses, two or three E subunit isoforms with molecular masses ranging from 29 to 40 kDa were detected in other plants. Two isotypes of V-ATPase from the pea epicotyl were separated by ion exchange chromatography and had subunit compositions differing only in the ratio of E1 and E2. There was a difference in the V(max) and K(m) of ATP hydrolysis between the two isotypes. E1 was scarcely detected in crude membrane fractions from the leaf and cotyledon, while E2 was detected in fractions from all of the tissues examined. The compositions of D subunit isoforms in the leaf and epicotyl were different, and the vacuolar membrane in the leaf did not contain D2. The efficiency of H(+) pumping activity in the vacuolar membrane of the leaf was higher than that of the epicotyl. The results suggest that the presence of the isoforms of D and E subunits is characteristic to plants and that the isoforms are closely related to the enzymatic properties.
Kawamura
Y
Y
Institute of Low Temperature Science, Hokkaido University, 060-0819 Sapporo, Japan.
Arakawa
K
K
Maeshima
M
M
Yoshida
S
S
D000595:Amino Acid Sequence; D002499:Centrifugation, Density Gradient; D015151:Immunoblotting; D007700:Kinetics; D008969:Molecular Sequence Data; D018532:Pisum sativum / Q000201:enzymology*; D010449:Peptide Mapping; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism; D020033:Protein Isoforms / Q000737:chemistry; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000378:metabolism*; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship; D025262:Vacuolar Proton-Translocating ATPases; D015003:Yeasts / Q000201:enzymology
D010940:Plant Proteins; D020033:Protein Isoforms; D025262:Vacuolar Proton-Translocating ATPases; D006180:Proton-Translocating ATPases