Alkaline phosphodiesterase I release from eucaryotic plasma membranes by phosphatidylinositol-specific phospholipase C. III. The release from tumor cells.
1. Alkaline phosphodiesterase I release from two tumor cell lines, KB III or AH-130 cells, by the action of phosphatidylinositol-specific phospholipase C (PIPLC) of Bacillus thuringiensis was studied. 2. A significant amount of alkaline phosphodiesterase I was released from both the cell suspension and homogenate of KB III cells, but not from AH-130 cells. 3. The release of the enzyme from KB III cells was dependent on, or proportional to, the reaction time and the PIPLC or cell concentrations. 4. Alkaline phosphatase and 5'-nucleotidase were also released from KB III cells, while gamma-glutamyl transpeptidase and dipeptidyl peptidase IV were not solubilized. The enzyme release by the action of PIPLC was suppressed when purified anti-PIPLC antibody was added to the reaction mixture. This suggests that the enzyme release must be due to the direct action of PIPLC on KB III cells. 5. The alkaline phosphodiesterase I released from KB III cells had a mol. wt of 240,000 and was activated by Mg2+, but strongly inhibited by EDTA and thiol reagents and by 5'-nucleotide-containing compounds. Although KB III cells were derived from Homo sapiens tumor, the released alkaline phosphodiesterase I appeared to be very similar to enzymes obtained from normal tissues of Rattus norvegicus.
Nakabayashi
T
T
Laboratory of Biochemistry, Faculty of Pharmaceutical Sciences, Mukogawa Women's University, Hyogo, Japan.
Matsuoka
Y
Y
Taguchi
R
R
Ikezawa
H
H
Nakane
H
H
Ono
K
K
Kimura
Y
Y
D015720:5'-Nucleotidase / Q000378:metabolism; D000818:Animals; D001413:Bacillus thuringiensis / Q000201:enzymology; D002462:Cell Membrane / Q000201:enzymology; D002850:Chromatography, Gel; D003470:Culture Media; D018819:Dipeptidyl Peptidase 4; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000378:metabolism; D005260:Female; D006801:Humans; D051379:Mice; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D043264:Phosphodiesterase I; D054801:Phosphoinositide Phospholipase C; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism*; D051381:Rats; D014407:Tumor Cells, Cultured; D005723:gamma-Glutamyltransferase / Q000378:metabolism
D003470:Culture Media; D005723:gamma-Glutamyltransferase; D015720:5'-Nucleotidase; D010727:Phosphoric Diester Hydrolases; D043264:Phosphodiesterase I; D054801:Phosphoinositide Phospholipase C; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; D018819:Dipeptidyl Peptidase 4; D043265:Phosphatidylinositol Diacylglycerol-Lyase