Glycosylation and processing of pro-B-type natriuretic peptide in cardiomyocytes

Biochem Biophys Res Commun. 2011 Aug 5;411(3):593-8. doi: 10.1016/j.bbrc.2011.06.192. Epub 2011 Jul 6.


B-type natriuretic peptide (BNP) and its related peptides are biomarkers for the diagnosis of heart failure. Recent studies identified several O-glycosylation sites, including Thr-71, on human pro-BNP but the functional significance was unclear. In this study, we analyzed glycosylation and proteolytic processing of pro-BNP in cardiomyocytes. Human pro-BNP wild-type (WT) and mutants were expressed in HEK 293 cells and murine HL-1 cardiomyocytes. Pro-BNP and BNP were analyzed by immunoprecipitation and Western blotting. Glycosidases and glycosylation inhibitors were used to examine carbohydrates on pro-BNP. The effects of furin and corin expression on pro-BNP processing in cells also were examined. We found that in HEK 293 cells, recombinant pro-BNP contained significant amounts of O-glycans with terminal oligosialic acids. Mutation at Thr-71 reduced O-glycans on pro-BNP and increased pro-BNP processing. In HL-1 cardiomyocytes, residue Thr-71 contained little O-glycans, and pro-BNP WT and T71A mutant were processed similarly. In HEK 293 cells, pro-BNP was processed by furin. Mutations at Arg-73 and Arg-76, but not Lys-79, prevented pro-BNP processing. In HL-1 cardiomyocytes, which express furin and corin, single or double mutations at Arg-73, Arg-76 and Lys-79 did not prevent pro-BNP processing. Only when all these three residues were mutated, was pro-BNP processing completely blocked. Our data indicate that pro-BNP glycosylation in cardiomyocytes differed significantly from that in HEK 293 cells. In HEK 293 cells, furin cleaved pro-BNP at Arg-76 whereas in cardiomyocytes corin cleaved pro-BNP at multiple residues including Arg-73, Arg-76 and Lys-79.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Furin / chemistry
  • Furin / metabolism
  • Glycoside Hydrolases / chemistry
  • Glycosylation
  • HEK293 Cells
  • Humans
  • Myocytes, Cardiac / metabolism*
  • Natriuretic Peptide, Brain / chemistry
  • Natriuretic Peptide, Brain / genetics
  • Natriuretic Peptide, Brain / metabolism*
  • Serine Endopeptidases / chemistry
  • Serine Endopeptidases / metabolism


  • Natriuretic Peptide, Brain
  • Glycoside Hydrolases
  • CORIN protein, human
  • Serine Endopeptidases
  • Furin