Journal Article

NECAP 1 Regulates AP-2 Interactions to Control Vesicle Size, Number, and Cargo During Clathrin-Mediated Endocytosis

Philie, Jacynthe, Murphy, Sebastian, Dokainish, Hatem, Gehring, Kalle, Halin, Marilene, McPherson, Peter S., Ritter, Brigitte, Gudheti, Manasa V., Kozlov, Guennadi, Girard, Martine, Jorgensen, Erik M.

2013

Overview

AP-2 is the core-organizing element in clathrin-mediated endocytosis. During the formation of clathrin-coated vesicles, clathrin and endocytic accessory proteins interact with AP-2 in a temporally and spatially controlled manner, yet it remains elusive as to how these interactions are regulated. Here, we demonstrate that the endocytic protein NECAP 1, which binds to the α-ear of AP-2 through a C-terminal WxxF motif, uses an N-terminal PH-like domain to compete with clathrin for access to the AP-2 β2-linker, revealing a means to allow AP-2-mediated coordination of accessory protein recruitment and clathrin polymerization at sites of vesicle formation. Knockdown and functional rescue studies demonstrate that through these interactions, NECAP 1 and AP-2 cooperate to increase the probability of clathrin-coated vesicle formation and to control the number, size, and cargo content of the vesicles. Together, our data demonstrate that NECAP 1 modulates the AP-2 interactome and reveal a new layer of organizational control within the endocytic machinery.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Adapter proteins / Adaptor Protein Complex 2 - metabolism / Amino Acid Sequence / Animals / Binding Sites / Binding sites (Biochemistry) / Binding, Competitive / Cercopithecus aethiops / Clathrin / Clathrin - metabolism / Clathrin-Coated Vesicles - metabolism / Conserved Sequence / COS Cells / Endocytosis / Experiments / Gene Knockdown Techniques / Health aspects / HEK293 Cells / Humans / Medical research / Membrane Proteins - chemistry / Membrane Proteins - metabolism / Models, Biological / Molecular Sequence Data / Physiological aspects / Protein Binding / Protein Structure, Tertiary / Protein Transport / Recruitment / Synaptic Vesicles - metabolism