Rapid purification of two lipocortin-like proteins from mice lung.
We have purified two proteins (40 kDa and 36 kDa) from mice lung by the method of calcium-precipitation/EGTA solubilization and then a separation on a high anion exchanger column (Mono Q HR 5/5. Pharmacia) with a gradient of NaCl. The two proteins were strong inhibitors of phospholipase A2 as assessed in vitro with porcine pancreatic phospholipase A2 and [3H]-oleic acid labeled E. Coli membranes as substrate. The 40 kDa protein had a pI of 5.8 and was found to be immunologically related to human recombinant lipocortin I. The 36 kDa protein had a pI of 4.7 and cross-reacted with a polyclonal antibody raised against a 32 kDa human lipocortin-like protein described in human blood mononuclear cells. We report here a rapid purification of two distinct lipocortin-like proteins from mice lung.
Errasfa
M
M
Unité associée INSERM n. 285/Institut Pasteur, Paris, France.
Russo-Marie
F
F
D000818:Animals; D017302:Annexins; D006023:Glycoproteins / Q000302:isolation & purification*; D007525:Isoelectric Focusing; D008168:Lung / Q000032:analysis*; D051379:Mice; D008970:Molecular Weight; D010741:Phospholipases A / Q000037:antagonists & inhibitors; D054467:Phospholipases A2; D011506:Proteins / Q000302:isolation & purification*
D017302:Annexins; D006023:Glycoproteins; D011506:Proteins; D010741:Phospholipases A; D054467:Phospholipases A2