Cargando…

The Cytosolic Phospholipase A(2)α N-Terminal C2 Domain Binds and Oligomerizes on Membranes with Positive Curvature

Group IV phospholipase A(2)α (cPLA(2)α) regulates the production of prostaglandins and leukotrienes via the formation of arachidonic acid from membrane phospholipids. The targeting and membrane binding of cPLA(2)α to the Golgi involves the N-terminal C2 domain, whereas the catalytic domain produces...

Descripción completa

Detalles Bibliográficos
Autores principales: Ward, Katherine E., Sengupta, Ranjan, Ropa, James P., Amiar, Souad, Stahelin, Robert V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7226022/
https://www.ncbi.nlm.nih.gov/pubmed/32331436
http://dx.doi.org/10.3390/biom10040647
_version_ 1783534190134820864
author Ward, Katherine E.
Sengupta, Ranjan
Ropa, James P.
Amiar, Souad
Stahelin, Robert V.
author_facet Ward, Katherine E.
Sengupta, Ranjan
Ropa, James P.
Amiar, Souad
Stahelin, Robert V.
author_sort Ward, Katherine E.
collection PubMed
description Group IV phospholipase A(2)α (cPLA(2)α) regulates the production of prostaglandins and leukotrienes via the formation of arachidonic acid from membrane phospholipids. The targeting and membrane binding of cPLA(2)α to the Golgi involves the N-terminal C2 domain, whereas the catalytic domain produces arachidonic acid. Although most studies of cPLA(2)α concern its catalytic activity, it is also linked to homeostatic processes involving the generation of vesicles that traffic material from the Golgi to the plasma membrane. Here we investigated how membrane curvature influences the homeostatic role of cPLA(2)α in vesicular trafficking. The cPLA(2)α C2 domain is known to induce changes in positive membrane curvature, a process which is dependent on cPLA(2)α membrane penetration. We showed that cPLA(2)α undergoes C2 domain-dependent oligomerization on membranes in vitro and in cells. We found that the association of the cPLA(2)α C2 domain with membranes is limited to membranes with positive curvature, and enhanced C2 domain oligomerization was observed on vesicles ~50 nm in diameter. We demonstrated that the cPLA(2)α C2 domain localizes to cholesterol enriched Golgi-derived vesicles independently of cPLA(2)α catalytic activity. Moreover, we demonstrate the C2 domain selectively localizes to lipid droplets whereas the full-length enzyme to a much lesser extent. Our results therefore provide novel insight into the molecular forces that mediate C2 domain-dependent membrane localization in vitro and in cells.
format Online
Article
Text
id pubmed-7226022
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-72260222020-05-18 The Cytosolic Phospholipase A(2)α N-Terminal C2 Domain Binds and Oligomerizes on Membranes with Positive Curvature Ward, Katherine E. Sengupta, Ranjan Ropa, James P. Amiar, Souad Stahelin, Robert V. Biomolecules Article Group IV phospholipase A(2)α (cPLA(2)α) regulates the production of prostaglandins and leukotrienes via the formation of arachidonic acid from membrane phospholipids. The targeting and membrane binding of cPLA(2)α to the Golgi involves the N-terminal C2 domain, whereas the catalytic domain produces arachidonic acid. Although most studies of cPLA(2)α concern its catalytic activity, it is also linked to homeostatic processes involving the generation of vesicles that traffic material from the Golgi to the plasma membrane. Here we investigated how membrane curvature influences the homeostatic role of cPLA(2)α in vesicular trafficking. The cPLA(2)α C2 domain is known to induce changes in positive membrane curvature, a process which is dependent on cPLA(2)α membrane penetration. We showed that cPLA(2)α undergoes C2 domain-dependent oligomerization on membranes in vitro and in cells. We found that the association of the cPLA(2)α C2 domain with membranes is limited to membranes with positive curvature, and enhanced C2 domain oligomerization was observed on vesicles ~50 nm in diameter. We demonstrated that the cPLA(2)α C2 domain localizes to cholesterol enriched Golgi-derived vesicles independently of cPLA(2)α catalytic activity. Moreover, we demonstrate the C2 domain selectively localizes to lipid droplets whereas the full-length enzyme to a much lesser extent. Our results therefore provide novel insight into the molecular forces that mediate C2 domain-dependent membrane localization in vitro and in cells. MDPI 2020-04-22 /pmc/articles/PMC7226022/ /pubmed/32331436 http://dx.doi.org/10.3390/biom10040647 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Ward, Katherine E.
Sengupta, Ranjan
Ropa, James P.
Amiar, Souad
Stahelin, Robert V.
The Cytosolic Phospholipase A(2)α N-Terminal C2 Domain Binds and Oligomerizes on Membranes with Positive Curvature
title The Cytosolic Phospholipase A(2)α N-Terminal C2 Domain Binds and Oligomerizes on Membranes with Positive Curvature
title_full The Cytosolic Phospholipase A(2)α N-Terminal C2 Domain Binds and Oligomerizes on Membranes with Positive Curvature
title_fullStr The Cytosolic Phospholipase A(2)α N-Terminal C2 Domain Binds and Oligomerizes on Membranes with Positive Curvature
title_full_unstemmed The Cytosolic Phospholipase A(2)α N-Terminal C2 Domain Binds and Oligomerizes on Membranes with Positive Curvature
title_short The Cytosolic Phospholipase A(2)α N-Terminal C2 Domain Binds and Oligomerizes on Membranes with Positive Curvature
title_sort cytosolic phospholipase a(2)α n-terminal c2 domain binds and oligomerizes on membranes with positive curvature
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7226022/
https://www.ncbi.nlm.nih.gov/pubmed/32331436
http://dx.doi.org/10.3390/biom10040647
work_keys_str_mv AT wardkatherinee thecytosolicphospholipasea2anterminalc2domainbindsandoligomerizesonmembraneswithpositivecurvature
AT senguptaranjan thecytosolicphospholipasea2anterminalc2domainbindsandoligomerizesonmembraneswithpositivecurvature
AT ropajamesp thecytosolicphospholipasea2anterminalc2domainbindsandoligomerizesonmembraneswithpositivecurvature
AT amiarsouad thecytosolicphospholipasea2anterminalc2domainbindsandoligomerizesonmembraneswithpositivecurvature
AT stahelinrobertv thecytosolicphospholipasea2anterminalc2domainbindsandoligomerizesonmembraneswithpositivecurvature
AT wardkatherinee cytosolicphospholipasea2anterminalc2domainbindsandoligomerizesonmembraneswithpositivecurvature
AT senguptaranjan cytosolicphospholipasea2anterminalc2domainbindsandoligomerizesonmembraneswithpositivecurvature
AT ropajamesp cytosolicphospholipasea2anterminalc2domainbindsandoligomerizesonmembraneswithpositivecurvature
AT amiarsouad cytosolicphospholipasea2anterminalc2domainbindsandoligomerizesonmembraneswithpositivecurvature
AT stahelinrobertv cytosolicphospholipasea2anterminalc2domainbindsandoligomerizesonmembraneswithpositivecurvature