TY  - JOUR
TI  - Purification of Cytosolic Phospholipase A2α C2-domain  after Expression in Soluble Form in Escherichia coli
AU  - Hirano, Yoshinori 
AU  - Gao, Yong Guang
AU  - Simanshu, Dhirendra K.
AU  - Stephenson, Daniel J.
AU  - Vu, Ngoc T.
AU  - Malinina, Lucy 
AU  - Chalfant, Charles E.
AU  - Pate, Dinshaw J.
AU  - Brown, Rhoderick E
VL  - 11
IS  - 3
PY  - 2021
DA  - 2021/02/05
SP  - e3906
C1  - Bio-protocol 2021;11:e3906
DO  - 10.21769/BioProtoc.3906
UR  - https://doi.org/10.21769/BioProtoc.3906
AB  - Previous expression/purification strategies for cytosolic phospholipase A2α C2-domain in Escherichia coli 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 A2α C2-domain in soluble form in E. coli 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 A2α (Hirano et al., 2019)
KW  - Cytosolic phospholipase A2α
KW  - C2-domain purification
KW  - Soluble expression in E. coli
KW  - 6xHis-SUMO tag
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False