Characterization of calcium-independent cytosolic phospholipase A2 activity in the submucosal regions of rat stomach and small intestine.
This study was undertaken to compare the calcium-independent phospholipase A2 (PLA2) activities in the cytosols of twelve rat tissues and to determine whether their activities were distinct. 1-O-Alk-1'-enyl-2-[14C]-oleoyl-sn-glycero-3-phosphocholine (PlsC) and 1-O-Alk-1'-enyl-2-[14C]oleoyl-sn-glycero-3-phosphoethanolamine (PlsE) were synthesized and used as substrates, instead of phosphatidyl compounds, to exclude hydrolysis by cytosolic PLA1 activity that could be present in some of the cytosolic preparations. For each tissue, we examined substrate specificity, pH optimum, and effect of adenosine triphosphate (ATP) and ATP analogues. PLA2 activity was detected in eleven out of the twelve issues examined. Based on substrate specificity and pH optimum, cytosolic calcium-independent PLA2 were classified in three groups. The first group, which included PLA2 from small intestine, stomach and spleen, had the highest specific activity with PlsC as substrate (1253, 309 and 75 nmol/mg protein/hour, respectively) and an optimal pH at 6.5. Activity with PlsE as substrate was much lower (20-37%) than with PlsC. The second group of PLA2 activities included the cytosolic activities from thymus, lung, liver and pancreas that showed lower specific activities for both substrates (14-23 nmol/mg protein/hour with PlsC) and had a broader optimal pH range of 6.1 to 7.5. The cytosols from brain, kidney, heart and muscle comprised the third PLA2 group that was found to have a higher specific activity with PlsE (5-20 nmol/mg protein/hour) than PlsC and an optimal pH range from 7.4 to 7.9.(ABSTRACT TRUNCATED AT 250 WORDS)
Fukushima
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W. Alton Jones Cell Science Center, Lake Placid, New York 12946.
Serrero
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D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D005591:Chemical Fractionation; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D003600:Cytosol / Q000201:enzymology*; D004228:Dithionitrobenzoic Acid / Q000494:pharmacology; D005753:Gastric Mucosa / Q000201:enzymology* / Q000648:ultrastructure; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000201:enzymology* / Q000648:ultrastructure; D007421:Intestine, Small / Q000201:enzymology / Q000648:ultrastructure; D008297:Male; D010741:Phospholipases A / Q000378:metabolism*; D054466:Phospholipases A1; D054467:Phospholipases A2; D010955:Plasmalogens / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013379:Substrate Specificity
D010955:Plasmalogens; D000255:Adenosine Triphosphate; D004228:Dithionitrobenzoic Acid; D010741:Phospholipases A; D054466:Phospholipases A1; D054467:Phospholipases A2; D002118:Calcium