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PMID: 25165141 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Flat clathrin lattices: stable features of the plasma membrane.

Molecular biology of the cell ·Vol. 25 ·No. 22 ·2014-11-05 ·Pages 3581-94

Grove J, Metcalf DJ, Knight AE, Wavre-Shapton ST, Sun T, Protonotarios ED, Griffin LD, Lippincott-Schwartz J, Marsh M

Abstract

Clathrin-mediated endocytosis (CME) is a fundamental property of eukaryotic cells. Classical CME proceeds via the formation of clathrin-coated pits (CCPs) at the plasma membrane, which invaginate to form clathrin-coated vesicles, a process that is well understood. However, clathrin also assembles into flat clathrin lattices (FCLs); these structures remain poorly described, and their contribution to cell biology is unclear. We used quantitative imaging to provide the first comprehensive description of FCLs and explore their influence on plasma membrane organization. Ultrastructural analysis by electron and superresolution microscopy revealed two discrete populations of clathrin structures. CCPs were typified by their sphericity, small size, and homogeneity. FCLs were planar, large, and heterogeneous and present on both the dorsal and ventral surfaces of cells. Live microscopy demonstrated that CCPs are short lived and culminate in a peak of dynamin recruitment, consistent with classical CME. In contrast, FCLs were long lived, with sustained association with dynamin. We investigated the biological relevance of FCLs using the chemokine receptor CCR5 as a model system. Agonist activation leads to sustained recruitment of CCR5 to FCLs. Quantitative molecular imaging indicated that FCLs partitioned receptors at the cell surface. Our observations suggest that FCLs provide stable platforms for the recruitment of endocytic cargo.

MeSH Terms
Animals CHO Cells Chemokine CCL5/metabolism,pharmacology Clathrin/metabolism Clathrin-Coated Vesicles/metabolism,ultrastructure Coated Pits, Cell-Membrane/metabolism,ultrastructure Cricetulus Dynamins/metabolism Endocytosis/physiology Genes, Reporter Green Fluorescent Proteins/metabolism HEK293 Cells HeLa Cells Humans Microscopy, Electron Microscopy, Fluorescence Molecular Imaging Protein Transport/drug effects Receptors, CCR5/agonists,metabolism
Chemicals
CCL5 protein, human CCR5 protein, human Chemokine CCL5 Clathrin Receptors, CCR5 Green Fluorescent Proteins Dynamins
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Grove Joe
MRC Laboratory for Molecular Cell Biology, London WC1E 6BT, United Kingdom Institute of Immunity and Transplantation, University College London, London NW3 2PF, United Kingdom j.grove@ucl.ac.uk m.marsh@ucl.ac.uk.
Metcalf Daniel J
Biophysics and Diagnostics, National Physical Laboratory, Teddington TW11 0LW, United Kingdom.
Knight Alex E
Biophysics and Diagnostics, National Physical Laboratory, Teddington TW11 0LW, United Kingdom.
Wavre-Shapton Silène T
MRC Laboratory for Molecular Cell Biology, London WC1E 6BT, United Kingdom.
Sun Tony
MRC Laboratory for Molecular Cell Biology, London WC1E 6BT, United Kingdom.
Protonotarios Emmanouil D
CoMPLEX, University College London, London WC1E 6BT, United Kingdom.
Griffin Lewis D
CoMPLEX, University College London, London WC1E 6BT, United Kingdom.
Lippincott-Schwartz Jennifer
Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892.
Marsh Mark
MRC Laboratory for Molecular Cell Biology, London WC1E 6BT, United Kingdom j.grove@ucl.ac.uk m.marsh@ucl.ac.uk.
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1939-4586
DOI
10.1091/mbc.E14-06-1154
Email
j.grove@ucl.ac.uk
m.marsh@ucl.ac.uk
Published
2014-11-05
Epub
2014-00-27
Pages
3581-94
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC4230618
Subset
IM
Grants
Medical Research Council · MC_UU_12018/1 · United Kingdom
Medical Research Council · MR/K015826/1 · United Kingdom
Medical Research Council · MC_U122665002 · United Kingdom
External Links
PubMed source DOI: 10.1091/mbc.E14-06-1154 PMC Full Text View journal details
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