Assaying phospholipase A2 activity.
نویسندگان
چکیده
Mammalian cells contain many structurally and functionally diverse phospholipases A2 (PLA2) that catalyze the hydrolysis of sn-2 fatty acid from membrane phospholipid. Assays are described for measuring the activity of Group IVA cytosolic PLA2alpha(cPLAalpha) and for secreted PLA2s (sPLA2) that are suitable for purified enzymes and for measuring activity in crude cell lysates and culture medium. The assay for cPLA2alpha involves measuring the calcium-dependent release of radiolabeled sn-2 arachidonic acid from small unilamellar vesicles of phosphatidylcholine. Methods are described for distinguishing cPLA2alpha activity in cell lysates from other PLA2s. sPLA2 activity is monitored using a fluorimetric assay that measures the continuous calcium-dependent formation of albumin-bound pyrene fatty acid from the sn-2 position of phosphatidylglycerol.
منابع مشابه
Effect of phospholipase A2 on temperature-induced high-affinity [3H]tryptamine binding sites in rat brain.
To investigate a link between membrane phospholipids and tryptamine binding molecules, we examined the effects of phospholipases A2 and D on the temperature-sensitive high-affinity [3H]tryptamine binding sites in rat brain. When the phospholipase A2-treated membranes were exposed to 1% bovine serum albumin (BSA) before assaying for [3H]tryptamine binding, a complete dose-dependent inhibition cu...
متن کاملAnti scelorotic Effect of Hydroalcoholic Extract of verbascum thapsus, Lemon Peel and Juice and Quinacrine on Blood Lipids, C-Reactive Protein and Phospholipase A2 Activity in Male Wistar Rats
Introduction: Atherosclerosis is a disease of the veins characterized by the deposition of low-density lipoprotein (LDL) cholesterol and total cholesterol on the inner wall of medium and large diameter arteries. The aim of this study was to compare the anti-sclerotic effect of hydroalcoholic extract of verbascum Thapsus, lemon juice, lemon peel and quinacrine on blood lipid concentration, C-rea...
متن کاملRaised serum activity of phospholipase A2 immunochemically related to group II enzyme in inflammatory bowel disease: its correlation with disease activity of Crohn's disease and ulcerative colitis.
Calcium dependent phospholipase A2 activity in the mixed micelles of 1-palmitoyl-2-oleoyl-phosphatidylglycerol and cholate was measured in sera of 39 patients with Crohn's disease, 40 patients with ulcerative colitis, and 40 healthy controls. The phospholipase A2 activity was significantly raised in those sera of the patients with active Crohn's disease and those with moderate and severe ulcera...
متن کاملSerum phospholipase A2 in patients after splenectomy.
Phospholipase A2 values increase in serum in various inflammatory states, infections, and postoperatively in surgical patients. Several organs, including the liver and spleen have been suggested as sources of circulating phospholipase A2. The purpose of the present work was to examine the possible role of the spleen as a source of elevated serum concentrations of phospholipase A2 after surgery....
متن کاملSerum phospholipases A2 after aortobifemoral reconstruction.
Phospholipase A2 has been implicated in the pathogenesis of local and distant tissue injury after ischaemia and reperfusion. A common operation inducing ischaemia and reperfusion is aortobifemoral reconstruction, during which the aorta is clamped and the blood supply via the inferior mesenteric artery and iliac arteries is interrupted. The purpose of the present work was to study the catalytic ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Methods in molecular biology
دوره 284 شماره
صفحات -
تاریخ انتشار 2004