Assay of phospholipases C and D in presence of other lipid hydrolases.
The activity of a phospholipase C or phospholipase D may be assessed by measuring the radioactivity or phosphate released into the aqueous phase of a lipid extract. However, in crude enzyme fractions, this type of analysis may not be possible due to formation of water-soluble metabolites by other enzymatic reactions, as demonstrated here with a crude lysosomal enzyme fraction. In such instances, analysis of both water-soluble and lipid-soluble metabolites, at various times of incubation, may still provide clear identification of phospholipases C or D, even when a variety of lipases and other hydrolases are present.
Hostetler
K Y
KY
Gardner
M F
MF
Aldern
K A
KA
D000818:Animals; D002250:Carbon Radioisotopes; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002855:Chromatography, Thin Layer / Q000379:methods; D006867:Hydrolases; D007202:Indicators and Reagents; D007700:Kinetics; D008055:Lipids / Q000302:isolation & purification; D008099:Liver / Q000201:enzymology*; D008247:Lysosomes / Q000201:enzymology*; D010713:Phosphatidylcholines; D010739:Phospholipase D / Q000032:analysis* / Q000378:metabolism; D010741:Phospholipases A / Q000032:analysis / Q000378:metabolism; D011865:Radioisotope Dilution Technique; D051381:Rats; D010738:Type C Phospholipases / Q000032:analysis* / Q000378:metabolism
D002250:Carbon Radioisotopes; D007202:Indicators and Reagents; D008055:Lipids; D010713:Phosphatidylcholines; D006867:Hydrolases; D010741:Phospholipases A; D010738:Type C Phospholipases; D010739:Phospholipase D