The glycosylphosphatidylinositol-anchored phosphatase from Spirodela oligorrhiza is a purple acid phosphatase.
We recently presented clear evidence that the major low-phosphate-inducible phosphatase of the duckweed Spirodela oligorrhiza is a glycosylphosphatidylinositol (GPI)-anchored protein, and, to our knowledge, is the first described from higher plants (N. Morita, H. Nakazato, H. Okuyama, Y. Kim, G.A. Thompson, Jr. [1996] Biochim Biophys Acta 1290: 53-62). In this report the purified 57-kD phosphatase is shown to be a purple metalloenzyme containing Fe and Mn atoms and having an absorption maximum at 556 nm. The phosphatase activity was only slightly inhibited by tartrate, as expected for a purple acid phosphatase (PAP). Furthermore, the protein cross-reacted with an anti-Arabidopsis PAP antibody on immunoblots. The N-terminal amino acid sequence of the phosphatase was very similar to those of Arabidopsis, red kidney bean (Phaseolus vulgaris), and soybean (Glycine max) PAP. Extracts of S. oligorrhiza plants incubated with the GPI-specific precursor [3H]ethanolamine were treated with antibodies raised against the purified S. oligorrhiza phosphatase. Radioactivity from the resulting immunoprecipitates was specifically associated with a 57-kD band on sodium dodecyl sulfate-polyacrylamide gels. These results, together with previous findings, strongly indicate that the GPI-anchored phosphatase of S. oligorrhiza is a PAP.
Nakazato
H
H
Laboratory of Environmental Molecular Biology, Graduate School of Environmental Earth Science, Hokkaido University, Kita-ku, Sapporo 060-0810, Japan.
Okamoto
T
T
Nishikoori
M
M
Washio
K
K
Morita
N
N
Haraguchi
K
K
Thompson
G A
GA
Okuyama
H
H
D000135:Acid Phosphatase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D000595:Amino Acid Sequence; D015153:Blotting, Western; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019856:Ethanolamine; D006023:Glycoproteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D008670:Metals / Q000032:analysis; D008969:Molecular Sequence Data; D010944:Plants / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid; D013057:Spectrum Analysis; D013644:Tartrates / Q000494:pharmacology; D014316:Tritium
D004791:Enzyme Inhibitors; D006023:Glycoproteins; D017261:Glycosylphosphatidylinositols; D008670:Metals; D013644:Tartrates; D014316:Tritium; D019856:Ethanolamine; C091137:purple acid phosphatase; D000135:Acid Phosphatase