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Heterogeneous distribution of PIP/sub 2/-specific phospholipase C in rat liver

Conference · · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
OSTI ID:6030191
; ;
Phosphatidylinositol 4,5-bisphosphate-specific phospholipase C (PIP/sub 2/-PLC) activity was found in both a plasma membrane-enriched fraction prepared by Percoll gradient centrifugation and cytosol isolated from rat liver. PIP/sub 2/-PLC was assayed in the presence of 50 mM Tris-HCl pH 7.4, 100 ..mu..M free Ca/sup 2 +/, 150 mM KCl, 0.1% deoxycholate, 50 ..mu..M (/sup 3/H-inositol)PIP/sub 2/. No PIP/sub 2/-PLC activity was released from the membrane fraction whe washed with 50 mM Tris-HCl, pH 7.4, under which conditions residual lactate dehydrogenase was completely removed. Approximately 70% of the membrane-associated activity was solubilized by 0.5 M KCl in 50 mM Tris-HCl pH 7.4 (S-PLC). 30% of the membrane-associated activity was especially resistant to solubilization (M-PLC); it remained in the particulate fraction following three successive washes with 0.5 M KCl and an extraction with 1% Triton X-100. Cytosolic PIP/sub 2/-PLC, S-PLC, and M-PLC hydrolyze PIP/sub 2/ at rates 4, 20, and 15-times greater than phosphatidylinositol and at rates 20, 430, and 60-times greater than phosphatidylcholine respectively. The three PIP/sub 2/-PLC activities are activated by calcium and enhanced by deoxycholate and monovalent salts.
Research Organization:
Brandeis Univ., Waltham, MA
OSTI ID:
6030191
Report Number(s):
CONF-870644-
Conference Information:
Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 46:6
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALKALINE EARTH METALS
ANIMALS
BODY
CALCIUM
CARBOXYLESTERASES
CELL CONSTITUENTS
CELL MEMBRANES
DIGESTIVE SYSTEM
DISTRIBUTION
ELEMENTS
ENZYME ACTIVITY
ENZYMES
ESTERASES
ESTERS
GLANDS
HYDROLASES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LIPASE
LIPIDS
LIVER
MAMMALS
MEMBRANES
METALS
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
ORGANS
PHOSPHOLIPIDS
RATS
RESPONSE MODIFYING FACTORS
RODENTS
SOLUBILITY
TISSUE DISTRIBUTION
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES