Prorenin activation and prohormone convertases in the mouse As4.1 cell line.
The precise identification of prorenin-processing enzymes has been hampered by the very low abundance of juxtaglomerular cells in the kidney. Recently, an immortalized renin-producing renal tumor cell line (As4.1) has been proposed as a model to carry out such studies. Despite the fact that they contain secretory granules, we found no evidence (on the basis of enzymatic assays of renin activity in the supernatant of the cells and of immunoprecipitations experiments) that the As4.1 cells can secrete active renin through the regulated pathway. As4.1 cells produce only renin-1, as they derive from a strain of mice expressing only one renin gene. However, stable transfection of these cells with a renin-2 expression plasmid increased the capacity of this cell line to secrete active renin in the regulated pathway. Northern blot and reverse transcriptase-polymerase chain reaction amplification (RT-PCR) assays revealed that furin, PACE4 and PC5 were the only members of the proprotein convertase (PC) family to be present in these cells. As PC5 is the only such enzyme with the demonstrated ability to process mouse prorenin 2, it may constitute a candidate enzyme for the processing of prorenin-2 in mouse juxtaglomerular cells. However, it is not likely to be involved in the processing of mouse prorenin 1.
Laframboise
M
M
Laboratory of Neurobiology and Vasoactive Peptides, Institut de Recherches Cliniques de Montréal, Québec, Canada.
Reudelhuber
T L
TL
Jutras
I
I
Brechler
V
V
Seidah
N G
NG
Day
R
R
Gross
K W
KW
Deschepper
C F
CF
D000818:Animals; D015152:Blotting, Northern; D002460:Cell Line / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D003080:Cold Temperature; D003594:Cytoplasmic Granules / Q000648:ultrastructure; D004792:Enzyme Precursors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D007668:Kidney / Q000166:cytology*; D051379:Mice; D008854:Microscopy, Electron; D010957:Plasmids; D011233:Precipitin Tests; D045706:Proprotein Convertase 2; D045702:Proprotein Convertase 5; D043484:Proprotein Convertases; D012333:RNA, Messenger / Q000032:analysis; D012083:Renin / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D013381:Subtilisins / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D019076:Transgenes / Q000502:physiology
D004792:Enzyme Precursors; D012333:RNA, Messenger; C513086:Pcsk6 protein, mouse; D045702:Proprotein Convertase 5; D043484:Proprotein Convertases; D012697:Serine Endopeptidases; D013381:Subtilisins; C473805:propeptide convertase 5, mouse; D045706:Proprotein Convertase 2; D012083:Renin