Structural basis of phosphatidylcholine recognition by the C2-domain of cytosolic phospholipase A2α

Abstract
Data availability
Article and author information
Metrics

Abstract

Ca²⁺-stimulated translocation of cytosolic phospholipase A₂α (cPLA₂α) to the Golgi induces arachidonic acid production, the rate-limiting step in pro-inflammatory eicosanoid synthesis. Structural insights into the cPLA₂α preference for phosphatidylcholine (PC)-enriched membranes have remained elusive. Here, we report cPLA₂α C2-domain structure (2.2Å resolution) containing bound 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC) and Ca²⁺ ions. Two Ca²⁺are complexed at locations previously reported for lipid-free C2-domain. One of these Ca²⁺along with a third Ca²⁺ bridge the C2-domain to the DHPC phosphate group, which also interacts with Asn65. Tyr96 plays a key role in lipid headgroup recognition via cation-π interaction with the PC trimethylammonium group. Mutagenesis analyses confirm Tyr96 and Asn65 function in PC binding selectivity by C2-domain and regulation of cPLA₂αactivity. The differing DHPC-binding mode of cPLA₂α C2-domain, compared to phosphatidylserine or phosphatidylinositol 4,5-bisphosphate binding by other C2-domains, expands and deepens knowledge of lipid-binding mechanisms mediated by C2-domains.

Data availability

Diffraction data have been deposited in PDB under the accession code 6IEJ

The following data sets were generated

1. Hirano Y
2. Cao Y-G
3. Stephenson DJ
4. Vu NT
5. Malinina L
6. Simanshu DK
7. Chalfont CE
8. Patel DJ
9. Brown RE
(2019) Structural basis of phosphatidylcholine recognition by the C2-domain of cytosolic phospholipase A2α
Protein Data Bank, 6IEJ.

http://www.rcsb.org/structure/6IEJ

Article and author information

Author details

Yoshinori Hirano

Structural Biology Program, Memorial Sloan Kettering Cancer Center, New York, United States

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-9888-1616
Yong-Guang Gao

Hormel Institute, University of Minnesota, Austin, United States

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-9359-4252
Daniel J Stephenson

Department of Cell Biology, Microbiology, and Molecular Biology, University of South Florida, Tampa, United States

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-5698-3400
Ngoc T Vu

Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, United States

Competing interests
The authors declare that no competing interests exist.
Lucy Malinina

Hormel Institute, University of Minnesota, Austin, United States

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-7973-1831
Dhirendra K Simanshu

Structural Biology Program, Memorial Sloan Kettering Cancer Center, New York, United States

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-9717-4618
Charles E Chalfant

Department of Cell Biology, Microbiology, and Molecular Biology, University of South Florida, Tampa, United States

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-5844-5235
Dinshaw J Patel

Structural Biology Program, Memorial Sloan Kettering Cancer Center, New York, United States

Competing interests
The authors declare that no competing interests exist.
Rhoderick E Brown

Hormel Institute, University of Minnesota, Austin, United States

For correspondence
reb@umn.edu

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-7337-3604

Funding

National Institutes of Health (HL125353)

Rhoderick E Brown

Ministry of Education, Culture, Sports, Science, and Technology

Yoshinori Hirano

U.S. Department of Veterans Affairs (I BX001792)

Charles E Chalfant

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.