Purification and characterization of phospholipase A2 inhibitory proteins from pig thyroid gland.
A 32 kDa phospholipase A2 inhibitory protein was isolated from pig thyroid gland after calcium precipitation and fast protein liquid anion-exchange chromatography. SDS-polyacrylamide gel electrophoresis revealed the purity of the protein. The protein activity was assessed by the inhibition of pancreatic phospholipase A2 on [3H]oleic acid-labelled Escherichia coli membranes as substrate and on the prostaglandin E2 production of cultured thyroid cells. The amino acid composition and the isoelectric point were quite similar to those of endonexin previously described in other tissues or cells. The cross-reactivity of a polyclonal antibody against a 32 kDa lipocortin from human peripheral blood mononuclear cells with our thyroidal 32 kDa protein confirmed its lipocortin nature. Before the purification by fast protein liquid chromatography, the Ca2+ pellet contained lipocortin I (35 kDa and its core protein 33 kDa) identified by its cross-reactivity with a polyclonal antibody.
Antonicelli
F
F
Laboratoire de Biochimie, UFR Sciences, Reims, France.
Rothhut
B
B
Martiny
L
L
Aguie-Aguie
G
G
Lambert
B
B
Bellon
G
G
Russo-Marie
F
F
Jacquemin
C
C
Haye
B
B
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D017302:Annexins; D002851:Chromatography, High Pressure Liquid; D015232:Dinoprostone; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000302:isolation & purification* / Q000494:pharmacology; D007118:Immunoassay; D008970:Molecular Weight; D010740:Phospholipases / Q000037:antagonists & inhibitors*; D010741:Phospholipases A / Q000037:antagonists & inhibitors*; D054467:Phospholipases A2; D011458:Prostaglandins E / Q000096:biosynthesis; D013552:Swine; D013961:Thyroid Gland / Q000032:analysis* / Q000187:drug effects / Q000378:metabolism
D000596:Amino Acids; D017302:Annexins; D006023:Glycoproteins; D011458:Prostaglandins E; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2; D015232:Dinoprostone