Furin-mediated processing of Pro-C-type natriuretic peptide.
C-type natriuretic peptide (CNP) is a member of the natriuretic peptide family that is involved in a variety of homeostatic processes. Here we characterize the processing essential for the conversion of the precursor, human pro-CNP, to the biologically active hormone. In human embryonic kidney 293 and chondrosarcoma SW 1353 cells, recombinant pro-CNP was converted into a mature peptide intracellularly as detected by Western analysis. Expression of recombinant human corin, a proatrial natriuretic peptide convertase, did not enhance the processing of pro-CNP in these cells. The processing of pro-CNP was inhibited in the presence of an inhibitor of the endoprotease furin but was not affected by inhibitors of matrix metalloproteinases and tumor necrosis factor-alpha convertase. In furin-deficient human colon adenocarcinoma LoVo cells, no conversion of recombinant pro-CNP to CNP was detected. Expression of recombinant human furin in LoVo cells restored the ability of these cells to process pro-CNP. Furthermore, incubation of purified recombinant human furin with LoVo cell lysate containing pro-CNP led to the conversion of the precursor to a mature peptide. The furin-processed CNP was shown to be biologically active in a cell-based cGMP assay. These results demonstrate that furin is a critical enzyme for the processing of human pro-CNP.
Department of Cardiovascular Research, Berlex Biosciences, Richmond, California 94804, USA.
Chengliang
C
Wu
Faye
F
Wu
Junliang
J
Pan
John
J
Morser
Qingyu
Q
Wu
D015153:Blotting, Western; D002460:Cell Line; D006152:Cyclic GMP / Q000378:metabolism; D045683:Furin; D005822:Genetic Vectors; D006801:Humans; D008954:Models, Biological; D020098:Natriuretic Peptide, C-Type / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry; D011498:Protein Precursors / Q000737:chemistry; D011994:Recombinant Proteins / Q000378:metabolism; D012697:Serine Endopeptidases / Q000378:metabolism; D013381:Subtilisins / Q000737:chemistry / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured
D010455:Peptides; D011498:Protein Precursors; D011994:Recombinant Proteins; D020098:Natriuretic Peptide, C-Type; C119812:CORIN protein, human; D012697:Serine Endopeptidases; D013381:Subtilisins; D045683:Furin; D006152:Cyclic GMP