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. 2021 Apr;17(4):383-386.
doi: 10.1038/s41589-021-00748-z. Epub 2021 Mar 1.

Orphan receptor GPR37L1 remains unliganded

Tony Ngo #  1 , Brendan P Wilkins #  2   3 , Sean S So #  2   3 , Peter Keov  2 , Kirti K Chahal  1 , Angela M Finch  4 , James L J Coleman  2 , Irina Kufareva  5 , Nicola J Smith  6   7
Affiliations

Orphan receptor GPR37L1 remains unliganded

Tony Ngo et al. Nat Chem Biol. 2021 Apr.
No abstract available

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Conflict of interest statement

Competing Interests

The authors declare no competing interests.

Figures

Figure 1:
Figure 1:. The corrected GPR37L1 construct does not have the apparent Gαs constitutive activity seen with p426-r37L1.
(a–b) Examination of the effects of p426-r37L1 on reporter signal in (a) CRE-luc and (b) CAMYEL assays. HEK293 cells were transiently co-transfected with the CRE-luc reporter or the CAMYEL BRET biosensor together with incorrect or corrected GPR37L1 construct or β2-adrenoceptor, as indicated. ‘Empty’ refers to empty plasmid DNA. For CRE-luc assay, data represent n=2 biological replicates each performed in triplicate on separate days; each data point is the average of the technical replicates for the corresponding biological replicate, and the bar height indicates the mean of biological replicates. For CAMYEL assay, data represent n=1 with technical replicates shown as dots. RLU, relative light units; FSK, forskolin. (c) Western blot for transient HEK293 cellular expression of r37L1, corrected GPR37L1 or controls, as indicated. Cerebellum from a male C57BL/6J mouse was used as a positive control. Dashed lane contained protein ladder only. Image is n=1. All raw data is available in the accompanying Source Data file.
Figure 2:
Figure 2:. The corrected GPR37L1 construct does not display constitutive or ligand-induced activity in any assays examined.
Examination of constitutive and ligand-induced cAMP elevation in HEK293 cells transfected with or without the corrected pcDNA3.1-GPR37L1, using (a) CRE-luc or (b) CAMYEL assays. β2-adrenoceptor +/− 10 μM salbutamol was used as a positive control for constitutive cAMP signaling. Final ligand concentrations were 10 μM. (a) n=4 biological replicates; (b) n=3 biological replicates except for controls, SHA68, head activator, bosentan, zibotentan, ambrisentan and TX14A, which were n=4. (c) CAMYEL assay performed as in (b) using CHO-K1 cells to control for cellular background. Data represents n=3 biological replicates. (d) Ligand-induced Gαs coupling was probed using the AP-TGFα-sheddase assay in stable FlpIN TREx HEK293 cells inducibly expressing GPR37L1-eYFP, transiently transfected with AP-TGFα and chimeric Gαq/s. β1 adrenoceptor stimulated with isoproterenol (ISO) was used as a positive control. Data represent n=3 biological replicates. (e–f) Constitutive suppression of FSK-stimulated cAMP was monitored using the CAMYEL BRET biosensor in HEK293T cells transiently co-transfected with CAMYEL and either the corrected GPR37L1 plasmid, Smoothened, or β2 adrenoceptor. Gαi3 was transiently co-transfected with GPR37L1 and Smoothened. Data represent n=4 biological replicates except for the β2 adrenoceptor (n=3). All data panels are presented as mean +/− standard deviation, where the bar height represents the average of all biological replicates each performed independently in three technical replicates. The averages of the technical triplicates for each biological replicate are shown as dots. All raw data is available in the accompanying Source Data file.
See this image and copyright information in PMC

References

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