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 Citations (Scopus)

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

Cytosolic Phospholipases A2
Phosphatidylcholines
Phosphorylcholine
Lipids
Ions
Mutagenesis
Eicosanoids
Phosphatidylserines
Phosphatidylinositols
Arachidonic Acid
Cations
Phosphates
C2 Domains
Membranes

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

Structural basis of phosphatidylcholine recognition by the C2-domain of cytosolic phospholipase A2α. / Hirano, Yoshinori; Gao, Yong-Guang; Stephenson, Daniel J.; Vu, Ngoc T.; Malinina, Lucy; Simanshu, Dhirendra K.; Chalfant, Charles E.; Patel, Dinshaw J.; Brown, Rhoderick E.

In: eLife, Vol. 8, e44760, 03.05.2019.

Research output: Contribution to journalArticle

Hirano, Y, Gao, Y-G, Stephenson, DJ, Vu, NT, Malinina, L, Simanshu, DK, Chalfant, CE, Patel, DJ & Brown, RE 2019, 'Structural basis of phosphatidylcholine recognition by the C2-domain of cytosolic phospholipase A2α', eLife, vol. 8, e44760. https://doi.org/10.7554/eLife.44760
Hirano, Yoshinori ; Gao, Yong-Guang ; Stephenson, Daniel J. ; Vu, Ngoc T. ; Malinina, Lucy ; Simanshu, Dhirendra K. ; Chalfant, Charles E. ; Patel, Dinshaw J. ; Brown, Rhoderick E. / Structural basis of phosphatidylcholine recognition by the C2-domain of cytosolic phospholipase A2α. In: eLife. 2019 ; Vol. 8.
@article{423a9cac77c34f05b71cabcc83e6b436,
title = "Structural basis of phosphatidylcholine recognition by the C2-domain of cytosolic phospholipase A2α",
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 {\AA}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.",
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",
author = "Yoshinori Hirano and Yong-Guang Gao and Stephenson, {Daniel J.} and Vu, {Ngoc T.} and Lucy Malinina and Simanshu, {Dhirendra K.} and Chalfant, {Charles E.} and Patel, {Dinshaw J.} and Brown, {Rhoderick E}",
year = "2019",
month = "5",
day = "3",
doi = "10.7554/eLife.44760",
language = "English (US)",
volume = "8",
journal = "eLife",
issn = "2050-084X",
publisher = "eLife Sciences Publications",

}

TY - JOUR

T1 - Structural basis of phosphatidylcholine recognition by the C2-domain of cytosolic phospholipase A2α

AU - Hirano, Yoshinori

AU - Gao, Yong-Guang

AU - Stephenson, Daniel J.

AU - Vu, Ngoc T.

AU - Malinina, Lucy

AU - Simanshu, Dhirendra K.

AU - Chalfant, Charles E.

AU - Patel, Dinshaw J.

AU - Brown, Rhoderick E

PY - 2019/5/3

Y1 - 2019/5/3

N2 - 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 Åresolution), 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.

AB - 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 Åresolution), 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.

KW - cPLA2 C2-domain lipid binding specificity

KW - structure of C2-domain complexed with dihexanoyl phosphatidylcholinePC

KW - FRET and SPR lipid binding analyses

KW - cPLA2 catatlytic activity analyses

KW - C2-domain membrane penetration

KW - lipid dipole potential

UR - http://www.scopus.com/inward/record.url?scp=85067371284&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85067371284&partnerID=8YFLogxK

U2 - 10.7554/eLife.44760

DO - 10.7554/eLife.44760

M3 - Article

C2 - 31050338

AN - SCOPUS:85067371284

VL - 8

JO - eLife

JF - eLife

SN - 2050-084X

M1 - e44760

ER -