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Purification and characterization of phospholipase A 2 inhibitory proteins from pig thyroid gland
Author(s) -
Antonicelli F.,
Rothhut B.,
Martiny L.,
Aguie-Aguie G.,
Lambert B.,
Bellon G.,
Russo-Marie F.,
Jacquemin C.,
Haye B.
Publication year - 1988
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(88)81273-0
Subject(s) - polyclonal antibodies , isoelectric point , biochemistry , microbiology and biotechnology , chemistry , ammonium sulfate precipitation , gel electrophoresis , antibody , biology , enzyme , size exclusion chromatography , immunology
A 32 kDa phospholipase A 2 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 A 2 on [ 3 H]oleic acid‐labelled Escherichia coli membranes as substrate and on the prostaglandin E 2 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 Ca 2+ pellet contained lipocortin I (35 kDa and its core protein 33 kDa) identified by its cross‐reactivity with a polyclonal antibody.