Purification and characterization of phospholipase A2 inhibitory proteins from pig thyroid gland

FEBS Letters

Antonicelli, F.²  Aguie-Aguie, G.²  Jacquemin, C.¹  Martiny, L.²  Bellon, G.⁴  Lambert, B.²  Haye, B.²  Rothhut, B.³  Russo-Marie, F.³ 

[1] U 96 INSERM, 78, rue du Général Leclerc, 94275 Le Kremlin-Bicětre Cédex, France;Laboratoire de Biochimie, UFR Sciences, BP 347, 51062 Reims Cédex France;Laboratoire de Biochimie, U 285 INSERM, Institut Pasteur, 25, rue du Docteur Roux, 75524 Paris Cédex 15 France;Laboratoire de Biochimie Médicale, UFR Médicine, UA 610 CNRS, 51062 Reims Cédex France

关键词: Lipocortin;    Endonexin;    Phospholipase A2;    Thyroid;    PLA2;    phospholipase A2;    PMSF;    phenylmethylsulfonyl fluoride;    Hepes;    N-2-hydroxyethylpiperazine-N-N′-2-ethanesulfonic acid;   

DOI  :  10.1016/0014-5793(88)81273-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown