抄録
The release of plasma-membrane-bound enzymes by phos-phatidylinositol-specific phospholipase C obtained from Bacillus thuringiensis was investigated. Among the ectoenzymes of plasma membrane tested, alkaline phosphodiesterase I was released markedly from rat kidney cortex slices, in addition to alkaline phosphatase and 5'-nucleotidase. Other membrane-bound enzymes; alanine aminopeptidase, leucine aminopeptidase, dipeptidyl peptid-ase, leucine aminopeptidase, dipeptidyl peptidase IV, esterase and γ-glutamyl transpeptidase could not be liberated from the treated slices.
Alkaline phosphodiesterase I was released linearly from rat kidney slices with the concentration of phosphatidylinositol-specific phospholipase C, but little enzyme was released from rat liver slices. Alkaline phosphodiesterase I separated from kidney tissue with n-butanol still retained phosphatidylinositol and was transformed into a lower molecular weight form by phosphatidylino-sitol-specific phospholipase C. This suggests an important function for phos-phatidylinositol in the binding of alkaline phosphodiesterase I to the plasma membrane of rat kidney cells.
The alkaline phosphodiesterase I released from rat kidney had a molecular weight of about 240, 000 and an isoelectric point (pI) of 5.4. The enzyme hydrolyzed the phosphodiester linkage of p-nitrophenyl-thymidine 5'-monophosphate at pH 8.9 and had a Km value of 0.3 mM. The enzyme was activated by Mg2+ and Ca2+, but was inhibited by EDTA. Strong inhibition took place on the addition of adenosine 5'-phosphosulfate or the nucleotide pyrophosphates, i.e., UDP-galactose and a, β-methylene ATP.
