Soluble form of the (pro)renin receptor generated by intracellular cleavage by furin is secreted in plasma.
The (pro)renin receptor [(P)RR] is a 35-kDa transmembrane protein that plays a pivotal role in angiotensin tissue generation and in nonproteolytic prorenin activation. We detected a soluble form of (P)RR [s(P)RR; 28 kDa] in the conditioned medium of cultured cells. The aims of our study were to identify the protease responsible for the generation of s(P)RR, the site of shedding, and to establish the existence of circulating s(P)RR in plasma. We identified furin as the protease responsible for the shedding of endogenous (P)RR based on the following: LoVo colon carcinoma cells devoid of active furin synthesize full-length (P)RR but do not secrete s(P)RR; transfection of Chinese hamster ovary cells with a plasmid coding for alpha1-antitrypsin Portland variant, an inhibitor of furin, completely inhibited the generation of s(P)RR, whereas addition of GM6001, an inhibitor of metalloproteases or of tumor necrosis factor-alpha protease inhibitor-1, an inhibitor of ADAM17, in the culture medium has no effect; when the cDNA coding for (P)RR was translated in vitro and incubated with recombinant furin or ADAM17, only furin was able to generate the 28 kDa-s(P)RR, and mutagenesis in the potential furin cleavage R275A/KT/R278A site abolished s(P)RR generation. Immunofluorescence study in glomerular epithelial cells showed that (P)RR was cleaved in the trans-Golgi, and coprecipitation experiments with renin showed that s(P)RR was present in plasma. In conclusion, our results show that s(P)RR is generated intracellularly by furin cleavage, and that s(P)RR detected in plasma is able to bind renin.
Cousin
Christelle
C
Institut National de la Santé et de la Recherche Médicale and Collège de France, Paris, France.
Bracquart
Diane
D
Contrepas
Aurelie
A
Corvol
Pierre
P
Muller
Laurent
L
Nguyen
Genevieve
G
D000704:Analysis of Variance; D000818:Animals; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D003593:Cytoplasm / Q000378:metabolism; D053595:Embryonic Stem Cells; D005455:Fluorescent Antibody Technique; D045683:Furin / Q000494:pharmacology*; D006801:Humans; D016133:Polymerase Chain Reaction; D011336:Probability; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D012083:Renin / Q000187:drug effects / Q000378:metabolism*; D012680:Sensitivity and Specificity; D012995:Solubility; D014162:Transfection; D039221:mRNA Cleavage and Polyadenylation Factors / Q000378:metabolism*
D012333:RNA, Messenger; D039221:mRNA Cleavage and Polyadenylation Factors; D045683:Furin; D012083:Renin