Studies of the phospholipase A2 activity of rat ileal mucosa

Abstract

A rapid and simple procedure has been used to determine phospholipase A2 activity (EC 3.1.1.4) in rat ileal mucosa. We used 14C-oleate-labeled Escherichia coli as substrate for the phospholipase activity and a 0.45-micron Millipore filter to separate the product of hydrolysis--the 14C-oleic acid--from the unhydrolyzed substrate. The phospholipase A2 activity was optimal at pH 9.8 and at 2 mM Ca2+, but another peak of activity appeared at pH 7.2. In addition, cell fractionation revealed yet another phospholipase A2 activity at pH 5.0 in the absence of Ca2+. These findings suggest the presence of more than one phospholipase A2 in the ileal mucosa and points to the possible use of a simple procedure for studying their distribution and properties.