Phosphorylation of the AP2 {micro} subunit by AAK1 mediates high affinity binding to membrane protein sorting signals

doi:10.1083/jcb.200111068

Published 4 March 2002. doi:10.1083/jcb.200111068

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© The Rockefeller University Press, 0021-9525/2002/3/791 $5.00
The Journal of Cell Biology, Volume 156, Number 5, March 4, 2002 791-795

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Phosphorylation of the AP2 µ subunit by AAK1 mediates high affinity binding to membrane protein sorting signals

Doris Ricotta¹, Sean D. Conner², Sandra L. Schmid², Kurt von Figura¹ and Stefan Höning¹

¹ Institute for Biochemistry II, University of Göttingen, 37073 Göttingen, Germany
² The Scripps Research Institute, La Jolla, CA 92037

Address correspondence to Stefan Höning, Institute for Biochemistry II, University of Göttingen, Heinrich-Duker-Weg 12, Göttingen 37073, Germany. Tel.: 49-551-392127. Fax.: 49-551-395979. E-mail: shoenin{at}gwdg.de

During receptor-mediated endocytosis, AP2 complexes act as abridge between the cargo membrane proteins and the clathrincoat by binding to sorting signals via the µ2 subunitand to clathrin via the ß subunit. Here we show thatbinding of AP2 to sorting signals in vitro is regulated by phosphorylationof the µ2 subunit of AP2. Phosphorylation of µ2enhances the binding affinity of AP2 for sorting motifs as muchas 25-fold compared with dephosphorylated AP2. The recognitionof sorting signals was not affected by the phosphorylation statusof the ${alpha}$ or ß2 subunit, suggesting that phosphorylationof µ2 is critical for regulation of AP2 binding to sortingsignals. Phosphorylation of µ2 occurs at a single threonineresidue (Thr-156) and is mediated by the newly discovered adaptor-associatedkinase, AAK1, which copurifies with AP2. We propose that phosphorylationof the AP2 µ2 subunit by AAK1 ensures high affinity bindingof AP2 to sorting signals of cargo membrane proteins duringthe initial steps of receptor-mediated endocytosis.

Key Words: clathrin; endocytosis; receptor; trafficking; coated vesicle

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