AP2M1 Antibody (Rabbit mAb) [M24D15] Datasheet

Biological Description

Specificity	AP2M1 Antibody (Rabbit mAb) [M24D15]detects endogenous levels of total AP2M1 protein.
Background	AP2M1 encodes the μ2 subunit of adaptor protein complex 2 (AP-2), the clathrin adaptor that selects cargo for clathrin-mediated endocytosis and synaptic vesicle recycling. The subunit is the part of the complex that reads sorting signals in the cytoplasmic tails of cell surface proteins. Many such proteins are marked for internalization by a short motif consisting of a tyrosine followed by two variable residues and then a bulky hydrophobic residue, and the μ2 subunit recognizes this motif through its C-terminal signal-binding domain. Crystal structures of that domain bound to signal peptides from the epidermal growth factor receptor and from the trans-Golgi network protein TGN38 show how the recognition works. Contrary to earlier expectation that the motif would form a tight turn, the bound peptides adopt an extended conformation, and specificity arises from separate hydrophobic pockets that accommodate the tyrosine and the hydrophobic leucine of the motif. In the crystals the domain forms dimers, which could strengthen and sharpen binding to dimeric receptors. Disruption of this machinery causes neurological disease. A recurrent de novo missense variant in AP2M1 was identified in individuals with developmental and epileptic encephalopathies whose phenotypic similarity exceeded chance expectation, and the same variant was found in further individuals with neurodevelopmental disorders and generalized epilepsy in a larger cohort. Modelling indicates that the substitution impairs conformational activation and the thermodynamic entropy of AP-2, and functional experiments in human cells and mouse astrocytes show impaired clathrin-mediated endocytosis of transferrin while protein stability, expression, membrane recruitment and localization remain intact. This establishes the variant as a recurrent pathogenic cause of developmental and epileptic encephalopathy and implicates early endocytic dysfunction in epilepsy.

Specificity

AP2M1 Antibody (Rabbit mAb) [M24D15]detects endogenous levels of total AP2M1 protein.

Background

AP2M1 encodes the μ2 subunit of adaptor protein complex 2 (AP-2), the clathrin adaptor that selects cargo for clathrin-mediated endocytosis and synaptic vesicle recycling. The subunit is the part of the complex that reads sorting signals in the cytoplasmic tails of cell surface proteins. Many such proteins are marked for internalization by a short motif consisting of a tyrosine followed by two variable residues and then a bulky hydrophobic residue, and the μ2 subunit recognizes this motif through its C-terminal signal-binding domain. Crystal structures of that domain bound to signal peptides from the epidermal growth factor receptor and from the trans-Golgi network protein TGN38 show how the recognition works. Contrary to earlier expectation that the motif would form a tight turn, the bound peptides adopt an extended conformation, and specificity arises from separate hydrophobic pockets that accommodate the tyrosine and the hydrophobic leucine of the motif. In the crystals the domain forms dimers, which could strengthen and sharpen binding to dimeric receptors. Disruption of this machinery causes neurological disease. A recurrent de novo missense variant in AP2M1 was identified in individuals with developmental and epileptic encephalopathies whose phenotypic similarity exceeded chance expectation, and the same variant was found in further individuals with neurodevelopmental disorders and generalized epilepsy in a larger cohort. Modelling indicates that the substitution impairs conformational activation and the thermodynamic entropy of AP-2, and functional experiments in human cells and mouse astrocytes show impaired clathrin-mediated endocytosis of transferrin while protein stability, expression, membrane recruitment and localization remain intact. This establishes the variant as a recurrent pathogenic cause of developmental and epileptic encephalopathy and implicates early endocytic dysfunction in epilepsy.

Usage Information

Application

WB

Dilution

WB

1:1000

Reactivity

Human, Mouse, Rat

Source

Rabbit Monoclonal Antibody

MW

50 kDa

Storage Buffer

PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage
(from the date of receipt)

-20°C (avoid freeze-thaw cycles), 2 years

References

https://pubmed.ncbi.nlm.nih.gov/31104773/
https://pubmed.ncbi.nlm.nih.gov/9812899/

AP2M1 Antibody (Rabbit mAb) [M24D15]

Biological Description

Usage Information

References

Application Data