Purification of Cytosolic Phospholipase Aα C2-domain after Expression in Soluble Form in .

Previous expression/purification strategies for cytosolic phospholipase Aα C2-domain in have relied on refolded protein recovered from inclusion bodies and sometimes containing C-terminal Cys139Ala and Cys141Ser substitutions to eliminate potential refolding complications induced by Cys residues. The protocol presented herein describes an effective method for the expression of cytosolic phospholipase Aα C2-domain in soluble form in and subsequent purification to homogeneity. This protocol, which utilizes a cleavable 6xHis-SUMO tag, has recently been used to gain insights into the structural basis of phosphatidylcholine recognition by the C2-domain of cytosolic phospholipase Aα ( Hirano , 2019 ).