EphA2 proteomics in human keratinocytes reveals a novel association with afadin and epidermal tight junctions.
J Cell Sci
; 130(1): 111-118, 2017 01 01.
Article
em En
| MEDLINE
| ID: mdl-27815408
ABSTRACT
EphA2 is a receptor tyrosine kinase that helps to maintain epidermal tissue homeostasis. A proximity-dependent biotin identification (BioID) approach was used to identify proteins in close proximity to EphA2 within primary human keratinocytes and three-dimensional (3D) reconstituted human epidermis (RHE) cultures to map a putative protein interaction network for this membrane receptor that exhibits a polarized distribution in stratified epithelia. Although a subset of known EphA2 interactors were identified in the BioID screen, >97% were uniquely detected in keratinocytes with over 50% of these vicinal proteins only present in 3D human epidermal culture. Afadin (AFDN), a cytoskeletal and junction-associated protein, was present in 2D and 3D keratinocyte cultures, and validated as a so-far-unknown EphA2-interacting protein. Loss of EphA2 protein disrupted the subcellular distribution of afadin and occludin in differentiated keratinocytes, leading to impairment of tight junctions. Collectively, these studies illustrate the use of the BioID approach in order to map receptor interaction networks in 3D human epithelial cultures, and reveal a positive regulatory role for EphA2 in the organization of afadin and epidermal tight junctions.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Queratinócitos
/
Junções Íntimas
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Receptor EphA2
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Proteômica
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Epiderme
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Proteínas dos Microfilamentos
Tipo de estudo:
Prognostic_studies
/
Risk_factors_studies
Limite:
Humans
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Male
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Newborn
Idioma:
En
Revista:
J Cell Sci
Ano de publicação:
2017
Tipo de documento:
Article
País de afiliação:
Estados Unidos