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

Yoshinori Hirano, Yong-Guang Gao, Daniel J. Stephenson, Ngoc T. Vu, Lucy Malinina, Dhirendra K. Simanshu, Charles E. Chalfant, Dinshaw J. Patel, Rhoderick E Brown

Research output: Contribution to journalArticle

2 Scopus citations

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 the structure of the cPLA2α C2-domain (at 2.2 Å
resolution), which contains bound 1,2-dihexanoyl-sn-glycero3-phosphocholine (DHPC) and Ca2+ ions. Two Ca2+ are complexed at previously reported locations in the lipid-free C2-domain. One of these Ca2+ions, along with a third Ca2+, bridges 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 that Tyr96 and Asn65 function in PC binding selectivity by the C2-domain and in the regulation of cPLA2α activity. The DHPC-binding mode of the cPLA2α C2-domain, which differs from phosphatidylserine or phosphatidylinositol 4,5-bisphosphate binding by other C2-domains, expands and deepens knowledge of the lipid-binding mechanisms mediated by C2-domains.
Original languageEnglish (US)
Article numbere44760
Number of pages28
JournaleLife
Volume8
DOIs
StatePublished - May 3 2019

    Fingerprint

Keywords

  • cPLA2 C2-domain lipid binding specificity
  • structure of C2-domain complexed with dihexanoyl phosphatidylcholinePC
  • FRET and SPR lipid binding analyses
  • cPLA2 catatlytic activity analyses
  • C2-domain membrane penetration
  • lipid dipole potential

PubMed: MeSH publication types

  • Journal Article
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

Cite this

Hirano, Y., Gao, Y-G., Stephenson, D. J., Vu, N. T., Malinina, L., Simanshu, D. K., Chalfant, C. E., Patel, D. J., & Brown, R. E. (2019). Structural basis of phosphatidylcholine recognition by the C2-domain of cytosolic phospholipase A2α. eLife, 8, [e44760]. https://doi.org/10.7554/eLife.44760