Purification and properties of lysophospholipase isoenzymes from pig gastric mucosa.
Two lysophospholipases, named gastric lysophospholipases I and II (enzymes I and II), were purified 3730- and 2680-fold from pig gastric mucosa. The preparations showed 22 and 23 kDa single protein bands on SDS/PAGE respectively. Both enzymes lacked transacylase activity and appeared to exist as monomers. Their activities were not affected by Ca2+, Mg2+ or EDTA. Enzyme I was most active at pH 8.5 and hydrolysed a variety of lysophospholipids including acidic lysophospholipids and the acyl analogue of platelet-activating factor, whereas enzyme II was most active at pH 8 and its activity was confined to lysophosphatidylcholine and lysophosphatidylethanolamine. When 1-palmitoylglycerophosphocholine was used as substrate, enzymes I and II showed half-maximal activities at 11 and 12 microM respectively. The enzymes exhibited no phospholipase B, lipase or general esterase activity. Enzyme II was significantly inhibited by lysophosphatidic acid whereas enzyme I was only moderately inhibited. Peptide mapping with V8 protease and papain revealed structural dissimilarity between the two enzymes. Antiserum raised against enzyme I did not recognize enzyme II, but did recognize the small-sized lysophospholipase purified from rat liver. Anti-(enzyme II) consistently did not cross-react with enzyme I or the liver enzyme. These antisera specifically recognized neither the 60 kDa lysophospholipase transacylase purified from liver nor any peritoneal macrophage protein. Thus gastric mucosa contains two different small-sized lysophospholipases: one is closely related to the small-sized lysophospholipase of liver, but the other appears to be a novel isoform.
Sunaga
H
H
Department of Biochemistry, Gunma University School of Medicine, Maebashi, Japan.
Sugimoto
H
H
Nagamachi
Y
Y
Yamashita
S
S
D000818:Animals; D015153:Blotting, Western; D005227:Fatty Acids / Q000494:pharmacology; D005753:Gastric Mucosa / Q000201:enzymology*; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism; D008245:Lysophospholipase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D008246:Lysophospholipids / Q000494:pharmacology; D008970:Molecular Weight; D010449:Peptide Mapping; D010712:Phosphatidic Acids / Q000494:pharmacology; D013379:Substrate Specificity; D013552:Swine
D005227:Fatty Acids; D007527:Isoenzymes; D008246:Lysophospholipids; D010712:Phosphatidic Acids; D008245:Lysophospholipase