TY - JOUR
T1 - A novel modelling mechanism of PAEL receptor and GABARAPL2 interaction involved in Parkinson's disease
AU - Dutta, Priyanka
AU - Dargahi, Leila
AU - O'Connell, Kara E.
AU - Bolia, Ashini
AU - Ozkan, Sefika
AU - Sailer, Andreas W.
AU - Dev, Kumlesh K.
N1 - Funding Information:
This work was supported in part by research grants from Trinity College Dublin Ireland , Novartis Pharma Basel Switerland , The Health Research Board Ireland and Science Foundation Ireland . We thank Davor Ivankovic, Debadutta Deb, Luke Healy, Louise Collins, Mirka Vanek, Fiona Crowe, Solene Fer and Eric Peterson for experimental support.
Publisher Copyright:
© 2018 Elsevier B.V.
PY - 2018/4/23
Y1 - 2018/4/23
N2 - Parkin associated endothelin like receptor (PAELR) is G-protein coupled and ubiquitinated by parkin, promoting its degradation. In autosomal recessive Parkinson's disease, mutations in parkin lead to PAELR aggregation in the endoplasmic reticulum (ER), ER stress, neurotoxicity and cell death. We have identified previously that the protein kinase C interacting protein (PICK1) interacts with and regulates the expression and cell toxicity of PAELR. Here, we experimentally identify and provide in-silico modelling of a novel interaction between PAELR and GABARAPL2 (γ-aminobutyrate type A receptor associated protein like 2), which is an autophagosome-specific Ub-like protein implicated in vesicle trafficking and autophagy. We show that the family of GABARAPs interact with the carboxy terminal (ct) of PAELR and find the cysteine rich region (-CCCCCC-EEC) of ct-PAELR interacts with the GABA A binding site of GABARAPL2. This interaction is modelled by in-slico analysis and confirmed using affinity chromatography, showing Myc-tagged GABARAPL2 is retained by a GST fusion of the ct-PAELR. We also demonstrate that transient transfection of GABARAPL2 in HEK293 cells reduces PAELR expression. This study supports the idea that protein levels of PAELR are likely regulated by a multitude of proteins including parkin, PICK1 and GABARAPL2 via mechanisms that include ubiquitination, proteasomal degradagtion and autophagy.
AB - Parkin associated endothelin like receptor (PAELR) is G-protein coupled and ubiquitinated by parkin, promoting its degradation. In autosomal recessive Parkinson's disease, mutations in parkin lead to PAELR aggregation in the endoplasmic reticulum (ER), ER stress, neurotoxicity and cell death. We have identified previously that the protein kinase C interacting protein (PICK1) interacts with and regulates the expression and cell toxicity of PAELR. Here, we experimentally identify and provide in-silico modelling of a novel interaction between PAELR and GABARAPL2 (γ-aminobutyrate type A receptor associated protein like 2), which is an autophagosome-specific Ub-like protein implicated in vesicle trafficking and autophagy. We show that the family of GABARAPs interact with the carboxy terminal (ct) of PAELR and find the cysteine rich region (-CCCCCC-EEC) of ct-PAELR interacts with the GABA A binding site of GABARAPL2. This interaction is modelled by in-slico analysis and confirmed using affinity chromatography, showing Myc-tagged GABARAPL2 is retained by a GST fusion of the ct-PAELR. We also demonstrate that transient transfection of GABARAPL2 in HEK293 cells reduces PAELR expression. This study supports the idea that protein levels of PAELR are likely regulated by a multitude of proteins including parkin, PICK1 and GABARAPL2 via mechanisms that include ubiquitination, proteasomal degradagtion and autophagy.
KW - GPR37
KW - In silico modelling
KW - Parkin-associated endothelial-like receptor (PAEL receptor)
KW - Parkinson's disease (PD)
KW - Protein-protein interaction
KW - γ-Aminobutyrate type a receptor associated protein like 2 (GABRAPL2)
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U2 - 10.1016/j.neulet.2018.02.055
DO - 10.1016/j.neulet.2018.02.055
M3 - Article
C2 - 29496607
AN - SCOPUS:85042676065
SN - 0304-3940
VL - 673
SP - 12
EP - 18
JO - Neuroscience Letters
JF - Neuroscience Letters
ER -