Structural basis of phosphatidylcholine recognition by the C2–domain of cytosolic phospholipase A2α
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 elusiv...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2019-05-01
|
| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/44760 |
| _version_ | 1828195630008238080 |
|---|---|
| author | 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 |
| author_facet | 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 |
| author_sort | Yoshinori Hirano |
| collection | DOAJ |
| description | 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-glycero-3-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. |
| first_indexed | 2024-04-12T09:46:52Z |
| format | Article |
| id | doaj.art-af6a1b2fae9e4f1fa435261f48886fe1 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T09:46:52Z |
| publishDate | 2019-05-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-af6a1b2fae9e4f1fa435261f48886fe12022-12-22T03:37:56ZengeLife Sciences Publications LtdeLife2050-084X2019-05-01810.7554/eLife.44760Structural basis of phosphatidylcholine recognition by the C2–domain of cytosolic phospholipase A2αYoshinori Hirano0https://orcid.org/0000-0001-9888-1616Yong-Guang Gao1https://orcid.org/0000-0002-9359-4252Daniel J Stephenson2https://orcid.org/0000-0002-5698-3400Ngoc T Vu3Lucy Malinina4https://orcid.org/0000-0001-7973-1831Dhirendra K Simanshu5https://orcid.org/0000-0002-9717-4618Charles E Chalfant6https://orcid.org/0000-0002-5844-5235Dinshaw J Patel7Rhoderick E Brown8https://orcid.org/0000-0002-7337-3604Structural Biology Program, Memorial Sloan-Kettering Cancer Center, New York, United States; Graduate School of Biological Sciences, Nara Institute of Science and Technology (NAIST), Takayama, JapanHormel Institute, University of Minnesota, Austin, United StatesDepartment of Biochemistry and Molecular Biology, Virginia Commonwealth University Medical Center, Richmond, United States; Department of Cell Biology, Microbiology and Molecular Biology, University of South Florida, Tampa, United StatesDepartment of Biochemistry and Molecular Biology, Virginia Commonwealth University Medical Center, Richmond, United StatesHormel Institute, University of Minnesota, Austin, United StatesStructural Biology Program, Memorial Sloan-Kettering Cancer Center, New York, United StatesDepartment of Cell Biology, Microbiology and Molecular Biology, University of South Florida, Tampa, United States; Research Service, James A. Haley Veterans Hospital, Tampa, United States; The Moffitt Cancer Center, Tampa, United StatesStructural Biology Program, Memorial Sloan-Kettering Cancer Center, New York, United StatesHormel Institute, University of Minnesota, Austin, United StatesCa2+-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-glycero-3-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.https://elifesciences.org/articles/44760C2-domain of cytoplasmic phospholipase A2 alphastructural mapping of phosphatidylcholine binding sitestructure-function analyses |
| spellingShingle | 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 Structural basis of phosphatidylcholine recognition by the C2–domain of cytosolic phospholipase A2α eLife C2-domain of cytoplasmic phospholipase A2 alpha structural mapping of phosphatidylcholine binding site structure-function analyses |
| title | Structural basis of phosphatidylcholine recognition by the C2–domain of cytosolic phospholipase A2α |
| title_full | Structural basis of phosphatidylcholine recognition by the C2–domain of cytosolic phospholipase A2α |
| title_fullStr | Structural basis of phosphatidylcholine recognition by the C2–domain of cytosolic phospholipase A2α |
| title_full_unstemmed | Structural basis of phosphatidylcholine recognition by the C2–domain of cytosolic phospholipase A2α |
| title_short | Structural basis of phosphatidylcholine recognition by the C2–domain of cytosolic phospholipase A2α |
| title_sort | structural basis of phosphatidylcholine recognition by the c2 domain of cytosolic phospholipase a2α |
| topic | C2-domain of cytoplasmic phospholipase A2 alpha structural mapping of phosphatidylcholine binding site structure-function analyses |
| url | https://elifesciences.org/articles/44760 |
| work_keys_str_mv | AT yoshinorihirano structuralbasisofphosphatidylcholinerecognitionbythec2domainofcytosolicphospholipasea2a AT yongguanggao structuralbasisofphosphatidylcholinerecognitionbythec2domainofcytosolicphospholipasea2a AT danieljstephenson structuralbasisofphosphatidylcholinerecognitionbythec2domainofcytosolicphospholipasea2a AT ngoctvu structuralbasisofphosphatidylcholinerecognitionbythec2domainofcytosolicphospholipasea2a AT lucymalinina structuralbasisofphosphatidylcholinerecognitionbythec2domainofcytosolicphospholipasea2a AT dhirendraksimanshu structuralbasisofphosphatidylcholinerecognitionbythec2domainofcytosolicphospholipasea2a AT charlesechalfant structuralbasisofphosphatidylcholinerecognitionbythec2domainofcytosolicphospholipasea2a AT dinshawjpatel structuralbasisofphosphatidylcholinerecognitionbythec2domainofcytosolicphospholipasea2a AT rhoderickebrown structuralbasisofphosphatidylcholinerecognitionbythec2domainofcytosolicphospholipasea2a |