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Article Abstract

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 ).

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7952920PMC
http://dx.doi.org/10.21769/BioProtoc.3906DOI Listing

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