GPR37 is processed in the N-terminal ectodomain by ADAM10 and furin
|Author:||Mattila, S. Orvokki1; Tuhkanen, Hanna E.1; Lackman, Jarkko J.1;|
1Medical Research Center Oulu, Research Unit of Biomedicine, University of Oulu, Oulu, Finland
2Unitat de Farmacologia, Departament de Patologia i Terapèutica Experimental, Facultat de Medicina i Ciències de la Salut, IDIBELL, Universitat de Barcelona, Barcelona, Spain
3Present address : Research Center and Memory Clinic, Fundació ACE, Institut Català de Neurociències Aplicades, International University of Catalunya (UIC), Barcelona, Spain
4Institute of Biochemistry, Kiel University, Kiel, Germany
5Institut de Neurociències, Universitat de Barcelona, Barcelona, Spain
|Online Access:||PDF Full Text (PDF, 1.6 MB)|
|Persistent link:|| http://urn.fi/urn:nbn:fi-fe2021060132573
John Wiley & Sons,
|Publish Date:|| 2021-06-01
GPR37 is an orphan G protein-coupled receptor (GPCR) implicated in several neurological diseases and important physiological pathways in the brain. We previously reported that its long N-terminal ectodomain undergoes constitutive metalloprotease-mediated cleavage and shedding, which have been rarely described for class A GPCRs. Here, we demonstrate that the protease that cleaves GPR37 at Glu167↓Gln168 is a disintegrin and metalloprotease 10 (ADAM10). This was achieved by employing selective inhibition, RNAi-mediated downregulation, and genetic depletion of ADAM10 in cultured cells as well as in vitro cleavage of the purified receptor with recombinant ADAM10. In addition, the cleavage was restored in ADAM10 knockout cells by overexpression of the wild type but not the inactive mutant ADAM10. Finally, postnatal conditional depletion of ADAM10 in mouse neuronal cells was found to reduce cleavage of the endogenous receptor in the brain cortex and hippocampus, confirming the physiological relevance of ADAM10 as a GPR37 sheddase. Additionally, we discovered that the receptor is subject to another cleavage step in cultured cells. Using site-directed mutagenesis, the site (Arg54↓Asp55) was localized to a highly conserved region at the distal end of the ectodomain that contains a recognition site for the proprotein convertase furin. The cleavage by furin was confirmed by using furin-deficient human colon carcinoma LoVo cells and proprotein convertase inhibitors. GPR37 is thus the first multispanning membrane protein that has been validated as an ADAM10 substrate and the first GPCR that is processed by both furin and ADAM10. The unconventional N-terminal processing may represent an important regulatory element for GPR37.
The FASEB journal
|Type of Publication:||
A1 Journal article – refereed
|Field of Science:||
1182 Biochemistry, cell and molecular biology
This work was supported by the Orion Research Foundation, Finnish Concordia Fund, Magnus Ehrnrooth Foundation (SOM), by travel grants from the Oskar Öflunds Stiftelse and FEBS (SOM), by Deutscher Forschungsgemeinschaft (SFB877-A3, PS), by FEDER/Ministerio de Ciencia, Innovación y Universidades–Agencia Estatal de Investigación (SAF2017-87349-R, FC), by the Catalan government (2017 SGR 1604, FC) and by the Academy of Finland (#295140, UEP-R.).
|Academy of Finland Grant Number:||
295140 (Academy of Finland Funding decision)
© 2021 The Authors. The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.