Occurrence of phospholipase A2 and lysophospholipase in a gastric H,K-ATPase-containing membrane fraction, and the formation of lysophosphatidylcholine in stimulated pig parietal cells.
A membrane fraction containing H,K-ATPase (EC 3.6.1.36) was prepared from pig gastric mucosa and found to contain phospholipase A2 (EC 3.1.1.4) and lysophospholipase (EC 3.1.1.5) activities. Washing the membranes decreased their protein content by 25%. Recovery profiles of H,K-ATPase, phospholipase A2 and lysophospholipase were similar for membranes washed either with water or with 0.15 or 1.5 M KCl. Nearly identical distribution profiles were obtained for the three enzyme activities after centrifugation of washed vesicle membranes on a linear sucrose gradient. The phospholipase A2 activity was stimulated by calcium and increased further in the presence of calmodulin. The amount of cellular radioactively labelled lysophosphatidylcholine was doubled upon cholinergic stimulation of isolated parietal cells prelabelled with [3H]glycerol or 32Pi. The liberated lyso[32P]phosphatidylcholine had its acyl chain in the sn-1 position, which implies an activation of a phospholipase A2. These findings indicate that secretagogues which increase the cytosolic Ca2+ concentration, i.e. acetylcholine, histamine and gastrin, may activate a phospholipase A2 in the parietal cell.
Olaisson
H
H
Department of Medical and Physiological Chemistry, University of Uppsala, Sweden.
Arvidson
G
G
Ma
J Y
JY
Mårdh
S
S
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000818:Animals; D005753:Gastric Mucosa / Q000166:cytology / Q000378:metabolism*; D017506:H(+)-K(+)-Exchanging ATPase; D066298:In Vitro Techniques; D008244:Lysophosphatidylcholines / Q000096:biosynthesis; D008245:Lysophospholipase / Q000378:metabolism*; D008566:Membranes / Q000201:enzymology; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D013552:Swine
D008244:Lysophosphatidylcholines; D010741:Phospholipases A; D054467:Phospholipases A2; D008245:Lysophospholipase; D000251:Adenosine Triphosphatases; D017506:H(+)-K(+)-Exchanging ATPase