Phosphorylation of threonine 156 of the μ2 subunit of the AP2 complex is essential for endocytosis in vitro and in vivo

TY - JOUR

T1 - Phosphorylation of threonine 156 of the μ2 subunit of the AP2 complex is essential for endocytosis in vitro and in vivo

AU - Olusanya, Oyinkan

AU - Andrews, Paul D.

AU - Swedlow, Jason R.

AU - Smythe, Elizabeth

N1 - Funding Information: We are very grateful to Alexander Sorkin (Denver) for the provision of the cDNA encoding HAμ2; to Margaret Robinson (Cambridge) for the provision of anti-μ2 and anti-clathrin antibodies; to Nick Morrice (MRC Protein Phosphorylation Unit, Dundee) for sequencing the phosphorylation site; and to Stefan Honing (Gottingen) for sharing unpublished results. We thank Colin Watts for helpful discussions and critical reading of the manuscript. This work was supported by the Medical Research Council (MRC) (Senior Fellowship to E.S.) and the Wellcome Trust (Career Development Award to J.R.S. and equipment grant for the purchase of the DeltaVision Restoration Microscope).

PY - 2001/6/5

Y1 - 2001/6/5

N2 - The clathrin-coated pit is the major port of entry for many receptors and pathogens and is the paradigm for membrane-based sorting events in higher cells [1]. Recently, it has been possible to reconstitute in vitro the events leading to assembly, invagination, and budding off of clathrin-coated vesicles, allowing dissection of the machinery required for sequestration of receptors into these structures [2-6]. The AP2 adaptor complex is a key element of this machinery linking receptors to the coat lattice, and it has previously been reported that AP2 can be phosphorylated both in vitro and in vivo [7-10]. However, the physiological significance of this has never been established. Here, we show that phosphorylation of a single threonine residue (Thr156) of the μ2 subunit of the AP2 complex is essential for efficient endocytosis of transferrin both in an in vitro coated-pit budding assay and in living cells.

AB - The clathrin-coated pit is the major port of entry for many receptors and pathogens and is the paradigm for membrane-based sorting events in higher cells [1]. Recently, it has been possible to reconstitute in vitro the events leading to assembly, invagination, and budding off of clathrin-coated vesicles, allowing dissection of the machinery required for sequestration of receptors into these structures [2-6]. The AP2 adaptor complex is a key element of this machinery linking receptors to the coat lattice, and it has previously been reported that AP2 can be phosphorylated both in vitro and in vivo [7-10]. However, the physiological significance of this has never been established. Here, we show that phosphorylation of a single threonine residue (Thr156) of the μ2 subunit of the AP2 complex is essential for efficient endocytosis of transferrin both in an in vitro coated-pit budding assay and in living cells.

UR - https://www.scopus.com/pages/publications/0035810915

U2 - 10.1016/S0960-9822(01)00240-8

DO - 10.1016/S0960-9822(01)00240-8

M3 - Article

C2 - 11516654

AN - SCOPUS:0035810915

SN - 0960-9822

VL - 11

SP - 896

EP - 900

JO - Current Biology

JF - Current Biology

IS - 11

ER -