Flat clathrin lattices: stable features of the plasma membrane.
Bottom Line: 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.
Affiliation: MRC Laboratory for Molecular Cell Biology, London WC1E 6BT, United Kingdom Institute of Immunity and Transplantation, University College London, London NW3 2PF, United Kingdom firstname.lastname@example.org email@example.com.Show MeSH
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Mentions: A number of reports suggest that FCLs may organize the cell surface distribution of both endocytic and nonendocytic receptors (Pumplin and Bloch, 1990; Miller et al., 1991; Sanan and Anderson, 1991; De Deyne et al., 1998; Lamaze et al., 2001; Kim et al., 2013). Therefore we evaluated the molecular codistribution of clathrin and CCR5 in our experimental system. To achieve this, we performed dual-color PALM/dSTORM superresolution imaging (Betzig et al., 2006; Bates et al., 2007; Heilemann et al., 2008) of photoactivatable, GFP-tagged clathrin light chain (PAGFP-LCa) and CCR5 labeled with mAb directly conjugated to Alexa Fluor 647. In untreated cells, CCR5 was distributed throughout the plasma membrane, organized into abundant puncta that likely represent individual or small clusters of receptors (Figure 6A). Critically, there was no ostensible association with FCLs. After 60 min of treatment with CCL5, the receptors had reorganized into larger clusters often arranged in and around FCLs, leaving the remaining plasma membrane predominantly devoid of receptor (Figure 6A).
Affiliation: MRC Laboratory for Molecular Cell Biology, London WC1E 6BT, United Kingdom Institute of Immunity and Transplantation, University College London, London NW3 2PF, United Kingdom firstname.lastname@example.org email@example.com.