Membrane-associated NAD+ glycohydrolase from rabbit erythrocytes is solubilized by phosphatidylinositol-specific phospholipase C.
NAD+ glycohydrolase (NADase) present on the surface of rabbit erythrocytes is a membrane-bound ectoenzyme that can be solubilized by phosphatidylinositol-specific phospholipase C (PIPLC). As much as 70% of the cell-associated NADase was made soluble by treatment with PIPLC. The portion of NADase that remained cell-associated after an initial PIPLC treatment proved to be resistant to subsequent solubilization attempts. Further analysis showed that release of NADase from erythrocytes could not be attributed to the action of proteinases or phospholipase C. Erythrocytes obtained from other mammals were analyzed and found to have variable amounts of PIPLC-susceptible NADase. Practically, this finding can be used to easily solubilize membrane-bound NADase as a first step in its purification.
Kim
U H
UH
Department of Biochemistry, Chonbuk National University Medical School, Chonju, Republic of Korea.
Rockwood
S F
SF
Kim
H R
HR
Daynes
R A
RA
D000818:Animals; D002712:Chlorides / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000201:enzymology*; D006801:Humans; D009244:NAD+ Nucleosidase / Q000097:blood*; D010618:Phenanthrolines / Q000494:pharmacology; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D010716:Phosphatidylinositols / Q000097:blood; D054801:Phosphoinositide Phospholipase C; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism* / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D011817:Rabbits; D012995:Solubility; D013045:Species Specificity; D015032:Zinc / Q000494:pharmacology; D017967:Zinc Compounds
D002712:Chlorides; D010618:Phenanthrolines; D010716:Phosphatidylinositols; D011480:Protease Inhibitors; D017967:Zinc Compounds; C016837:zinc chloride; D010727:Phosphoric Diester Hydrolases; D054801:Phosphoinositide Phospholipase C; D009244:NAD+ Nucleosidase; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D015032:Zinc; C025205:1,10-phenanthroline