| 1988 |
AP50 (AP2M1) was molecularly cloned and identified as the 50 kDa medium-chain protein component of the clathrin-coated vesicle coat assembly protein complex AP-2, encoding a 435 amino acid polypeptide of 49,612 Da that is highly conserved between rat and bovine brain. |
Molecular cloning and amino acid sequencing from rat brain cDNA libraries |
DNA (Mary Ann Liebert, Inc.) |
High |
3148444
|
| 1993 |
AP50 (AP2M1) is phosphorylated in vivo and in vitro at a single site, Thr-156, by an AP50 kinase activity that co-purifies with the AP complex (AP-1 and a soluble ~280 kDa protein complex); AP50 itself cannot autophosphorylate. |
In vitro phosphorylation, tryptic peptide mapping, automated Edman degradation, synthetic peptide substrate assay, gel filtration |
The Biochemical journal |
High |
8257432
|
| 1995 |
The human CLAPM1 (AP2M1) gene maps to chromosome region 3q28 by chromosomal in situ hybridization. |
Chromosomal fluorescence in situ hybridization (FISH) of a human genomic clone |
Genomics |
Medium |
8595912
|
| 1997 |
AP2M1 (AP50), the medium chain of AP-2, directly binds the cytoplasmic domain of CTLA-4 via the YXXΦ motif (GVYVKM, residues 199–204); mutation of Y201 abrogates binding and causes accumulation of CTLA-4 at the cell surface, demonstrating that AP2M1 mediates ligand-independent endocytosis of CTLA-4 into clathrin-coated vesicles. |
Yeast two-hybrid, co-immunoprecipitation, cell surface expression assay, site-directed mutagenesis (Y201 mutation) |
Journal of immunology |
High |
9200449 9256472
|
| 1997 |
Phosphorylation of CTLA-4 Y201 abolishes its binding to AP2M1 (AP50) and instead enables binding to the p85 subunit of PI3K (and weakly to SHP-2/SHP-1), demonstrating that the phosphorylation status of the YXXΦ motif determines which downstream effector is recruited. |
Yeast two-hybrid, co-immunoprecipitation in 293T cells, CTLA-4 phosphopeptide vs. unphosphorylated peptide binding assays |
Proceedings of the National Academy of Sciences of the United States of America |
High |
9200449 9256472
|
| 1999 |
The Drosophila ortholog of AP2M1 (AP50) is 86% identical to mouse and human AP2M1, is encoded by a single-copy gene at polytene chromosome position 94B1-B2, and is highly expressed in the central nervous system and midgut caecum during embryogenesis, consistent with conserved function in clathrin-mediated endocytosis in neurons. |
EST database identification, cDNA cloning, polytene chromosome mapping, in situ hybridization |
Gene |
Medium |
10375633
|
| 2003 |
AP2M1 (AP50) binds the cytoplasmic domain of B cell co-receptor CD22 via tyrosine-based internalization motifs; Tyr843 constitutes the primary binding site, and either Tyr843 or Tyr863 is sufficient for mAb-mediated internalization of CD22 via the AP-2 complex. |
Yeast two-hybrid, co-immunoprecipitation (co-precipitation of alpha-adaptin), transfectant Jurkat cell internalization assays with wild-type and mutant CD22 |
Journal of immunology |
High |
12646615
|
| 2008 |
AP2M1 (AP50) binds the alpha1b-adrenergic receptor (alpha1b-AR) via a YXXΦ motif in its C-tail; the alpha1a-AR does not bind AP50. Phosphorylation sites in the alpha1b-AR C-tail are required for beta-arrestin interaction (the dominant endocytic trigger), while AP50 binding alone is insufficient to drive efficient internalization. |
Co-immunoprecipitation, beta-arrestin translocation assays, biotinylation experiments, confocal microscopy, RNA interference |
Molecular pharmacology |
High |
18523139
|
| 2012 |
A conserved YXXΦ motif in HCV core protein mediates direct binding to AP2M1; this interaction recruits AP2M1 to lipid droplets, promotes core trafficking away from lipid droplets toward the trans-Golgi network, and is essential for viral assembly but not RNA replication. AAK1 and GAK kinases, which phosphorylate AP2M1 at Thr156, stimulate core–AP2M1 binding and are required for HCV assembly. |
Microfluidics affinity analysis, protein-fragment complementation assay, co-immunoprecipitation in infected cells, YXXΦ mutagenesis, AP2M1 siRNA knockdown, dominant-negative AP2M1 overexpression, quantitative confocal immunofluorescence, pharmacological kinase inhibition |
PLoS pathogens |
High |
22916011
|
| 2018 |
AP2M1 is identified as a downstream target of the cdk4–EZH2 axis during chemotherapy-induced senescence; AP2M1 is involved in transmission of secreted signals from senescent cells (likely via receptor endocytosis), and its upregulation by EZH2 facilitates escape from senescence in colorectal/breast cancer cells. |
Quantitative proteomic analysis, siRNA knockdown of EZH2, pharmacological EZH2 inhibition, cell emergence assay |
Cell death & disease |
Medium |
29415991
|
| 2019 |
A recurrent de novo p.Arg170Trp variant in AP2M1 impairs the conformational activation of the AP-2 complex and significantly reduces clathrin-mediated endocytosis of transferrin in human cells and in astrocytes from AP-2μ conditional knockout mice, without affecting AP2M1 stability, expression, membrane recruitment, or localization. |
Whole-exome sequencing, protein dynamics modeling, functional complementation of p.Arg170Trp in human cells, transferrin endocytosis assay in AP-2μ conditional KO mouse astrocytes |
American journal of human genetics |
High |
31104773
|
| 2020 |
AP2M1 is exploited by multiple viruses (influenza A, ZIKV, HIV, MERS-CoV, SARS-CoV-2, enterovirus A71) through their conserved YXXΦ motifs; AP2M1 depletion or YXXΦ mutation causes incorrect localization of viral proteins (e.g., failure of IAV nucleoprotein nuclear import, loss of ER localization of ZIKV-NS3), suppressing viral replication. The small molecule ACA disrupts AP2M1–virus interactions and inhibits viral replication in vitro and in vivo. |
YxxΦ mutagenesis of viral proteins, AP2M1 siRNA depletion, pharmacological inhibition (ACA compound), immunofluorescence localization, in vitro and in vivo antiviral assays across multiple virus types |
Science advances |
High |
32923629
|
| 2021 |
AP2M1 mediates autophagy-induced clathrin-dependent endocytosis and lysosomal degradation of the tight junction protein CLDN2 (claudin-2). AP2M1 binds CLDN2 via YXXΦ motifs (residues 67–70 and 148–151), and increased AP2M1 phosphorylation upon starvation promotes CLDN2-LC3 interaction. AP2M1 knockout prevents autophagy-induced CLDN2 degradation and enhances intestinal TJ permeability. |
Co-immunoprecipitation, immunolocalization, membrane fractionation, pharmacological inhibition of clathrin-mediated endocytosis, site-directed mutagenesis of CLDN2 YXXΦ motifs, AP2M1 CRISPR knockout, in vitro (cell lines), in vivo (mouse colon), ex vivo (human colon) |
Autophagy |
High |
34964704
|
| 2021 |
AP2M1 (dpy-23 in C. elegans) supports TGF-β signaling for collagen expression by limiting caveolin-1 (CAV-1) expression; loss of dpy-23/AP2M1 upregulates cav-1 in hypodermis, which increases clathrin-independent endocytosis and reduces TGF-β receptor I (TβRI) levels, thereby reducing TGF-β signaling and collagen production. |
C. elegans suppressor screen, RNA-seq, cav-1 RNAi knockdown, genetic epistasis (dpy-23 × lon-2 double mutant) |
International journal of molecular sciences |
Medium |
33561975
|
| 2021 |
AP2M1 interacts with the planar cell polarity protein Vangl2 at both its N-terminus and C-terminal Prickle-binding domain; knockdown of AP2M1 in developing cortical neurons reduces dendritic branching similarly to Vangl2 knockdown, establishing AP2M1-mediated endocytosis as necessary for dendrite morphogenesis. |
Yeast two-hybrid screen from mouse brain lysate, pull-down assay, shRNA knockdown in cortical neurons with dendritic branching quantification |
Genes to cells |
Medium |
34626136
|
| 2021 |
LRRK2 directly binds and phosphorylates AP2M1 (μ2 subunit of AP-2); loss of LRRK2 reduces AP2M1 phosphorylation (required for initial clathrin-coated vesicle formation), while overexpression or gain-of-function LRRK2 G2019S inhibits CCV uncoating at later stages, blocking new CCV formation cycles. LRRK2-dependent AP2M1 phosphorylation is brain-specific and mediates dopaminergic neurodegeneration in a Drosophila PD model. |
Kinase binding and phosphorylation assays, analysis of SH-SY5Y cells, mouse neurons and tissues (LRRK2 KO and G2019S knockin), Drosophila genetic model of PD with neurodegeneration readout |
Science signaling |
High |
34315807
|
| 2025 |
In a Drosophila model of AP2M1-DEE, pan-neuronal RNAi knockdown of the AP-2μ ortholog causes heat-sensitive paralysis and altered class IV dendritic arborization neuron morphology; a CRISPR-engineered p.Arg170Trp fly shows a milder seizure-resistant phenotype, suggesting the epilepsy in AP2M1-DEE may arise from broader neuronal developmental defects rather than direct synaptic dysfunction. |
Pan-neuronal RNAi, CRISPR knock-in of p.Arg170Trp, Drosophila behavioral assays (heat-sensitive paralysis, electrically induced seizures), dendritic morphology analysis |
Disease models & mechanisms |
Medium |
41017589
|
| 2026 |
In APP-overexpressing neurons, GluA2 (AMPAR subunit) undergoes enhanced endocytosis driven by increased interaction with AP2M1; a competitive peptide (G2CT) targeting the GluA2–AP2M1 interaction restores GluA2 membrane expression, improves synaptic function in vivo, and rescues cognitive deficits in 5×FAD mice without altering amyloid processing. |
Co-immunoprecipitation, surface biotinylation, competitive peptide (G2CT) treatment, in vivo electrophysiology, behavioral cognitive testing in 5×FAD mice, human AD brain tissue validation |
Neuropharmacology |
High |
41740904
|
| 2026 |
NAK family kinases (AAK1, GAK, BMP2K) phosphorylate AP2M1 at Thr156, activating AP2M1 and promoting its interaction with the YXXΦ motif in PRRSV glycoprotein GP5 and the receptor CD163, which is critical for efficient PRRSV trafficking to early endosomes; disruption of this phosphorylation or blockade of the AP2M1–YXXΦ interaction significantly impairs PRRSV internalization. |
Genetic (siRNA/CRISPR) and pharmacological inhibition of AAK1/GAK/BMP2K, AP2M1 phosphorylation site mutagenesis (Thr156), co-immunoprecipitation, viral entry/trafficking assays |
Transboundary and emerging diseases |
High |
41982333
|