Figure 2.
Cleavage of pro-ANP depended on the catalytic activity of corin. (a) Pro-ANP expression vector pcDNAproANP was cotransfected into 293 cells with vectors expressing wild-type corin (pcDNACorin, lane 2) and mutant corin S985A (pcDNACorin S985A, lane 1) or a control vector (pcDNA, lane 3). Pro-ANP and its derivatives were detected by Western blotting using an anti-V5 antibody. (b) Expression of recombinant corin in transfected 293 cells. To verify expression of recombinant corin in transfected cells, cell lysate was prepared and analyzed by SDS/PAGE and Western blotting. Expression of recombinant corin was detected in pcDNACorinV5 and pcDNACorin S985A but not pcDNA, transfected cells. A nonspecific (ns) band was present in samples from pcDNACorinV5, pcDNACorinS985A, and pcDNA transfected 293 cells.