Purification of cytosolic phospholipase A(2)α C2-domain after expression in soluble form in Escherichia coli Journal Article

   


Authors: Hirano, Y.; Gao, Y. G.; Simanshu, D. K.; Stephenson, D. J.; Vu, N. T.; Malinina, L.; Chalfant, C. E.; Patel, D. J.; Brown, R. E.
Article Title: Purification of cytosolic phospholipase A(2)α C2-domain after expression in soluble form in Escherichia coli
Abstract: 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). © 2021 Bio-protocol LLC. All Rights Reserved.
Keywords: c2-domain purification; 6xhis- sumo tag; cytosolic phospholipase a2α; soluble expression in e. coli
Journal Title: Bio-protocol
Volume: 11
Issue: 3
ISSN: 2331-8325
Publisher: Bio-protocol  
Date Published: 2021-02-01
Start Page: e3906
Language: English
DOI: 10.21769/BioProtoc.3906
PROVIDER: scopus
PMCID: PMC7952920
PUBMED: 33732793
DOI/URL:

https://www.scopus.com/inward/record.uri?eid=2-s2.0-85116287699&doi=10.21769%2fBioProtoc.3906&partnerID=40&md5=cedb864f7e5d05a5363ed8083e1f9cb3
Notes: Article -- Source: Scopus
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MSK Authors
  1. Dinshaw J Patel
    477 Patel
  2. Dhirendra Kumar Simanshu
    26 Simanshu
  3. Yoshinori Hirano
    2 Hirano
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