title: The AP-2 complex is excluded from the dynamic population of plasma membrane-associated clathrin.
abstract: Numerous biologically relevant substrates are selectively internalized via clathrin-mediated endocytosis. At the plasma membrane the AP-2 complex plays a major role in clathrin coat formation, interacting with both cargo and clathrin. Utilizing simultaneous dual-channel total internal reflection fluorescence microscopy we have analyzed components of the AP-2 complex (alpha- and beta 2-adaptin) during clathrin-mediated endocytosis. Although in static images enhanced green fluorescent protein-tagged AP-2 markers significantly co-localized with clathrin and other components of clathrin-coated pits, AP-2 did not seem to be present in clathrin spots that appeared to undergo internalization or motility parallel to the plane of the plasma membrane. Two populations of clathrin at the plasma membrane seem to exist, the dynamic and the static, and AP-2 appears to be only found within the latter. These results suggest that colocalized clathrin/AP-2 puncta may represent loci for coated pit production and that previous models that assumed AP-2 was retained within clathrin coats during endocytosis may need to be re-evaluated.
authors.affiliation: The Laboratory of Cellular Biophysics, The Rockefeller University, New York, New York 10021, USA.
authors.forename: Joshua Z
authors.initials: JZ
authors.lastname: Rappoport
authors.forename: Bushra W
authors.initials: BW
authors.lastname: Taha
authors.forename: Simone
authors.initials: S
authors.lastname: Lemeer
authors.forename: Alexandre
authors.initials: A
authors.lastname: Benmerah
authors.forename: Sanford M
authors.initials: SM
authors.lastname: Simon
mesh_terms: D033962:Adaptor Protein Complex 2 / Q000737:chemistry / Q000502:physiology*; D033965:Adaptor Protein Complex alpha Subunits / Q000737:chemistry; D033966:Adaptor Protein Complex beta Subunits / Q000737:chemistry; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D002966:Clathrin / Q000737:chemistry / Q000378:metabolism*; D018076:DNA, Complementary / Q000378:metabolism; D004705:Endocytosis; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D008077:Lipoproteins, LDL / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000378:metabolism
chemical_list: D033962:Adaptor Protein Complex 2; D033965:Adaptor Protein Complex alpha Subunits; D033966:Adaptor Protein Complex beta Subunits; D002966:Clathrin; D018076:DNA, Complementary; D008077:Lipoproteins, LDL