Phospholipases A2 of rod outer segment-free bovine retinae are different from well-known phospholipases A2.
We have recently demonstrated the presence of phospholipase A2 (PLA2) activity in a rod outer segment-free retinal fraction which we called P200 and which contains neuronal cells, Müller cells and rod inner segments. We report here our results on the characterization of this P200-PLA2 activity. We show that P200 probably contains more than one type of PLA2, as indicated by the results obtained with different chromatographically eluted PLA2-active fractions which were treated with either Ca2+, EGTA, dithiothreitol (DTT) or p-bromophenacyl bromide (pBPB), or heated. Moreover, the results from PLA2 assays using different substrates, as well as those obtained after treatment of the homogenate with H2SO4, guanosine 5'-O-(3-thio)triphosphate (GTPgammaS) and ATP, suggest that P200-PLA2 are different from well-known secretory PLA2, cytosolic PLA2 and Ca2+-independent PLA2. Control experiments using our 'back-and-forth'-thin layer chromatography (bf-TLC) technique allowed us to confirm that, in our assay conditions, the release of fatty acids was due to PLA2 enzymes. These results, which constitute the first characterization of PLA2 of the neural retina, thus suggest that it contains novel types of PLA2 enzyme, in contrast to well-known PLA2.
Jacob
M
M
GREIB, Département de Chimie-Biologie, Université du Québec à Trois-Rivières, 3351 Boul. des Forges, C.P. 500, Trois-Rivières, Québec, Canada.
Weech
P K
PK
Salesse
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D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D002852:Chromatography, Ion Exchange; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005227:Fatty Acids / Q000378:metabolism; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D066298:In Vitro Techniques; D007700:Kinetics; D009474:Neurons / Q000201:enzymology; D010741:Phospholipases A / Q000302:isolation & purification / Q000378:metabolism*; D054467:Phospholipases A2; D012160:Retina / Q000166:cytology / Q000201:enzymology*; D017948:Retinal Rod Photoreceptor Cells / Q000201:enzymology; D012374:Rod Cell Outer Segment / Q000201:enzymology; D013379:Substrate Specificity; D013464:Sulfuric Acids / Q000494:pharmacology
D004791:Enzyme Inhibitors; D005227:Fatty Acids; D013464:Sulfuric Acids; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D000255:Adenosine Triphosphate; D010741:Phospholipases A; D054467:Phospholipases A2; C033158:sulfuric acid; D002118:Calcium