Ectoenzyme release from rat liver and kidney by phosphatidylinositol-specific phospholipase C.
Ectoenzyme release from rat liver and kidney by phosphatidylinositol (PI)-specific phospholipase C of Bacillus thuringiensis was studied. Alkaline phosphatase and 5'-nucleotidase were released from rat kidney slices to extents of up to 60% and 30%, respectively. Release of alkaline phosphatase was observed at lower amounts of PI-specific phospholipase C than that of 5'-nucleotidase. Both enzymes were more easily released from microsomal fractions or free cells. From kidney cells, alkaline phosphatase was released without cell lysis, and more than 80% release of alkaline phosphatase was observed at 3.8% hydrolysis of PI. Isoelectric focusing profiles of alkaline phosphatase released by PI-specific phospholipase C were significantly different from the control in the cases of both rat liver and kidney. Lubrol-solubilized alkaline phosphatase was eluted at the void volume of a Toyopearl HW-55 column, while the enzyme obtained by further treatment with PI-specific phospholipase C was eluted in the lower-molecular-weight region corresponding to 100,000-110,000 daltons. Furthermore, Lubrol-solubilized phosphatase became more thermostable on treatment with PI-specific phospholipase C.
Taguchi
R
R
Asahi
Y
Y
Ikezawa
H
H
D015720:5'-Nucleotidase; D000469:Alkaline Phosphatase / Q000378:metabolism*; D000818:Animals; D001413:Bacillus thuringiensis / Q000201:enzymology; D002845:Chromatography; D006358:Hot Temperature; D007525:Isoelectric Focusing; D007668:Kidney / Q000201:enzymology*; D008099:Liver / Q000201:enzymology*; D008297:Male; D008861:Microsomes / Q000201:enzymology; D009708:Nucleotidases / Q000378:metabolism*; D010716:Phosphatidylinositols / Q000378:metabolism; D010740:Phospholipases / Q000502:physiology*; D051381:Rats; D011919:Rats, Inbred Strains; D012510:Sarcoma 180 / Q000201:enzymology; D013379:Substrate Specificity; D010738:Type C Phospholipases / Q000502:physiology*
D010716:Phosphatidylinositols; D010740:Phospholipases; D009708:Nucleotidases; D000469:Alkaline Phosphatase; D015720:5'-Nucleotidase; D010738:Type C Phospholipases