Apr 18, 2020

BRET sensors unravel that Plasmodium falciparum serpentine receptor 12 (PfSR12) increases surface expression of mammalian GPCRs in HEK293 cells

BioRxiv : the Preprint Server for Biology
P. H. S. PereiraC. R. Garcia

Abstract

Malaria causes millions of deaths worldwide and is considered a huge public health problem for underdeveloped countries. The most severe cases of malaria present complications of the host circulatory system, which may cause clogging and rupture of blood vessels, leading to death or important sequelae. Because of the previously suggested role of thrombin and platelet aggregation in Plasmodium falciparum biology, we hypothesized that one of the GPCR-like proteins identified in the genome of the parasite, P. falciparum serpentine receptor 12 (PfSR12), could be a thrombin-activated GPCR. To test this hypothesis we used a series of Bioluminescence and Bioluminescence Resonance Energy Transfer (BRET)-based biosensors to investigate the signaling activity of PfSR12. Using an Obelin based biosensor, thrombin promoted a PfSR12-dependent cytosolic Ca2+ rise in HEK293 cells. This Ca2+ mobilization was accompanied by DAG formation and PKC activation as detected using DAG and PKC BRET-based biosensors indicating a Gq/PLC/IP3 signaling pathway. The role of Gq was confirm using Gq/11 knockout HEK293 cells as well as the Gq-selective inhibitor, YM254890. Further investigation revealed that PfSR12 is not itself a thrombin receptor but rather pr...Continue Reading

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Mentioned in this Paper

Genome
Gene Expression
Genomics
Seattle Cancer Care Alliance
Analysis
Psychological Correlates
Single Nucleotide Polymorphism
Software Tools

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