Clathrin-mediated endocytosis (CME) is key to maintaining the transmembrane protein composition of cells' limiting membranes. During mammalian CME, a reversible phosphorylation event occurs on Thr156 of the μ2 subunit of the main endocytic clathrin adaptor, AP2. We show that this phosphorylation event starts during clathrin-coated pit (CCP) initiation and increases throughout CCP lifetime. μ2Thr156 phosphorylation favors a new, cargo-bound conformation of AP2 and simultaneously creates a binding platform for the endocytic NECAP proteins but without significantly altering AP2's cargo affinity in vitro. We describe the structural bases of both. NECAP arrival at CCPs parallels that of clathrin and increases with μ2Thr156 phosphorylation. In turn, NECAP recruits drivers of late stages of CCP formation, including SNX9, via a site distinct from where NECAP binds AP2. Disruption of the different modules of this phosphorylation-based temporal regulatory system results in CCP maturation being delayed and/or stalled, hence impairing global rates of CME.

• Klíčová slova
• AAK1, AP2 endocytic adaptor, NECAP, NMR, Numb-associated kinases (NAK), SNX9, TIRF, clathrin-mediated endocytosis, crystallography, regulation by phosphorylation,
• MeSH
• adaptorový proteinový komplex - alfa-podjednotky genetika   MeSH
• adaptorový proteinový komplex 2 genetika   metabolismus   MeSH
• endocytóza genetika   MeSH
• fosforylace genetika   MeSH
• klathrin genetika   metabolismus   MeSH
• klathrinové vezikuly genetika   metabolismus   MeSH
• lidé MeSH
• potažené jamky v buněčné membráně genetika   metabolismus   MeSH
• třídící nexiny genetika   MeSH
• vazba proteinů genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adaptorový proteinový komplex - alfa-podjednotky MeSH
• adaptorový proteinový komplex 2 MeSH
• klathrin MeSH
• NECAP1 protein, human MeSH Prohlížeč
• SNX9 protein, human MeSH Prohlížeč
• třídící nexiny MeSH