Rapid purification of calcium‐activated protease by calcium‐dependent hydrophobic‐interaction chromatography

FEBS Letters

Head, James F.¹  Gopalakrishna, Rayudu¹ 

[1] Department of Physiology, Boston University School of Medicine, 80 East Concord St., Boston, MA 02118, USA

关键词: Calpain;    Ca2+-activated protease;    Hydrophobic-interaction chromatography;    Leupeptin;    Phenyl-Sepharose;   

DOI  :  10.1016/0014-5793(85)80717-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Both low Ca²⁺- and high Ca²⁺-requiring forms of Ca²⁺-activated protease (calpains I and II) were found to bind to phenyl-Sepharose in a calcium-dependent manner, suggesting that both enzymes expose a hydrophobic surface region in the presence of Ca²⁺. Inclusion of leupeptin in column buffers prevented the loss of activity during hydrophobic-interaction and substrate-affinity chromatography. Under these conditions calpain II (high calcium-requiring form) was rapidly purified from bovine brain and rabbit skeletal muscle using successive phenyl-Sepharose and casein-Sepharose columns.

【 授权许可】

Unknown