Molecular composition and ultrastructure of the caveolar coat complex.

Caveolae are an abundant feature of the plasma membrane of many mammalian cell types, and have key roles in mechano-transduction, metabolic regulation, and vascular permeability. Caveolin and cavin proteins, as well as EHD2 and pacsin 2, are all present in caveolae. How these proteins assemble to fo...

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Autores principales:	Alexander Ludwig, Gillian Howard, Carolina Mendoza-Topaz, Thomas Deerinck, Mason Mackey, Sara Sandin, Mark H Ellisman, Benjamin J Nichols
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/9466bd7b95cc4bbcbb34ce0705f121b0
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spelling	oai:doaj.org-article:9466bd7b95cc4bbcbb34ce0705f121b02021-11-18T05:37:52ZMolecular composition and ultrastructure of the caveolar coat complex.1544-91731545-788510.1371/journal.pbio.1001640https://doaj.org/article/9466bd7b95cc4bbcbb34ce0705f121b02013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24013648/?tool=EBIhttps://doaj.org/toc/1544-9173https://doaj.org/toc/1545-7885Caveolae are an abundant feature of the plasma membrane of many mammalian cell types, and have key roles in mechano-transduction, metabolic regulation, and vascular permeability. Caveolin and cavin proteins, as well as EHD2 and pacsin 2, are all present in caveolae. How these proteins assemble to form a protein interaction network for caveolar morphogenesis is not known. Using in vivo crosslinking, velocity gradient centrifugation, immuno-isolation, and tandem mass spectrometry, we determine that cavins and caveolins assemble into a homogenous 80S complex, which we term the caveolar coat complex. There are no further abundant components within this complex, and the complex excludes EHD2 and pacsin 2. Cavin 1 forms trimers and interacts with caveolin 1 with a molar ratio of about 1∶4. Cavins 2 and 3 compete for binding sites within the overall coat complex, and form distinct subcomplexes with cavin 1. The core interactions between caveolin 1 and cavin 1 are independent of cavin 2, cavin 3, and EHD2 expression, and the cavins themselves can still interact in the absence of caveolin 1. Using immuno-electron microscopy as well as a recently developed protein tag for electron microscopy (MiniSOG), we demonstrate that caveolar coat complexes form a distinct coat all around the caveolar bulb. In contrast, and consistent with our biochemical data, EHD2 defines a different domain at the caveolar neck. 3D electron tomograms of the caveolar coat, labeled using cavin-MiniSOG, show that the caveolar coat is composed of repeating units of a unitary caveolar coat complex.Alexander LudwigGillian HowardCarolina Mendoza-TopazThomas DeerinckMason MackeySara SandinMark H EllismanBenjamin J NicholsPublic Library of Science (PLoS)articleBiology (General)QH301-705.5ENPLoS Biology, Vol 11, Iss 8, p e1001640 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Biology (General) QH301-705.5
spellingShingle	Biology (General) QH301-705.5 Alexander Ludwig Gillian Howard Carolina Mendoza-Topaz Thomas Deerinck Mason Mackey Sara Sandin Mark H Ellisman Benjamin J Nichols Molecular composition and ultrastructure of the caveolar coat complex.
description	Caveolae are an abundant feature of the plasma membrane of many mammalian cell types, and have key roles in mechano-transduction, metabolic regulation, and vascular permeability. Caveolin and cavin proteins, as well as EHD2 and pacsin 2, are all present in caveolae. How these proteins assemble to form a protein interaction network for caveolar morphogenesis is not known. Using in vivo crosslinking, velocity gradient centrifugation, immuno-isolation, and tandem mass spectrometry, we determine that cavins and caveolins assemble into a homogenous 80S complex, which we term the caveolar coat complex. There are no further abundant components within this complex, and the complex excludes EHD2 and pacsin 2. Cavin 1 forms trimers and interacts with caveolin 1 with a molar ratio of about 1∶4. Cavins 2 and 3 compete for binding sites within the overall coat complex, and form distinct subcomplexes with cavin 1. The core interactions between caveolin 1 and cavin 1 are independent of cavin 2, cavin 3, and EHD2 expression, and the cavins themselves can still interact in the absence of caveolin 1. Using immuno-electron microscopy as well as a recently developed protein tag for electron microscopy (MiniSOG), we demonstrate that caveolar coat complexes form a distinct coat all around the caveolar bulb. In contrast, and consistent with our biochemical data, EHD2 defines a different domain at the caveolar neck. 3D electron tomograms of the caveolar coat, labeled using cavin-MiniSOG, show that the caveolar coat is composed of repeating units of a unitary caveolar coat complex.
format	article
author	Alexander Ludwig Gillian Howard Carolina Mendoza-Topaz Thomas Deerinck Mason Mackey Sara Sandin Mark H Ellisman Benjamin J Nichols
author_facet	Alexander Ludwig Gillian Howard Carolina Mendoza-Topaz Thomas Deerinck Mason Mackey Sara Sandin Mark H Ellisman Benjamin J Nichols
author_sort	Alexander Ludwig
title	Molecular composition and ultrastructure of the caveolar coat complex.
title_short	Molecular composition and ultrastructure of the caveolar coat complex.
title_full	Molecular composition and ultrastructure of the caveolar coat complex.
title_fullStr	Molecular composition and ultrastructure of the caveolar coat complex.
title_full_unstemmed	Molecular composition and ultrastructure of the caveolar coat complex.
title_sort	molecular composition and ultrastructure of the caveolar coat complex.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/9466bd7b95cc4bbcbb34ce0705f121b0
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Molecular composition and ultrastructure of the caveolar coat complex.

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