Physical evidence for the presence of two forms of phosphatidate phosphohydrolase in rat liver.
Phosphatidate phosphohydrolase (PAP)-catalysed dephosphorylation of phosphatidic acid to diacylglycerol is an important step in glycerolipid metabolism and cell-signalling. Gel filtration chromatography on Superose 6 and anion-exchange chromatography on Mono Q of rat liver subcellular fractions has provided physical evidence for the presence of two distinct forms of PAP activity. One form was sensitive to inhibition by N-ethylmaleimide (NEM), had an apparent M(r) of 540,000 and was eluted from the anion-exchange column by 0.35 M NaCl, while the other was insensitive to inhibition by NEM, had an apparent M(r) of 240,000 and was eluted from the anion-exchange column by 0.15 M NaCl. Studies on the subcellular distribution of these two enzymes, using 5'-nucleotidase as a plasma membrane marker, demonstrated that the NEM-sensitive form was predominantly cytosolic but translocated to the microsomal membranes in response to oleate. The NEM-insensitive form was predominantly located in the plasma membrane but a small proportion (approx. 10%) of total cell activity was present on the endoplasmic reticulum. The implications of these results for the likely roles of the two different forms of PAP in fatty acid esterification and cell-signalling are discussed.
Day
C P
CP
Department of Biochemistry, Medical School, University of Newcastle, Newcastle upon Tyne, UK.
Yeaman
S J
SJ
D015720:5'-Nucleotidase / Q000378:metabolism; D000818:Animals; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D005033:Ethylmaleimide / Q000494:pharmacology; D007527:Isoenzymes / Q000032:analysis* / Q000037:antagonists & inhibitors; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D008297:Male; D010711:Phosphatidate Phosphatase / Q000032:analysis* / Q000037:antagonists & inhibitors; D051381:Rats; D011919:Rats, Inbred Strains; D013347:Subcellular Fractions / Q000201:enzymology
D007527:Isoenzymes; D010711:Phosphatidate Phosphatase; D015720:5'-Nucleotidase; D005033:Ethylmaleimide