Characterization of recombinant human HBP/CAP37/azurocidin, a pleiotropic mediator of inflammation-enhancing LPS-induced cytokine release from monocytes.
Neutrophil-derived heparin-binding protein (HBP) is a strong chemoattractant for monocytes. We report here for the first time the expression of recombinant HBP. A baculovirus containing the human HBP cDNA mediated in insect cells the secretion of a 7-residue N-terminally extended HBP form (pro-HBP). Deletion of the pro-peptide-encoding cDNA sequence resulted in correctly processed HBP at the N-terminus. Electrospray mass spectrum analysis of recombinant HBP yielded a molecular weight of 27.237 +/- 3 amu. Consistent with this mass is a HBP form of 225 amino acids (mature part +3 amino acid C-terminal extension). The biological activity of recombinant HBP was confirmed by its chemotactic action towards monocytes. Furthermore, we have shown that recombinant HBP stimulates in a dose-dependent manner the lipopolysaccharide (LPS)-induced cytokine release from human monocytes.
Rasmussen
P B
PB
Health Care Discovery, Novo Nordisk, Bagsvaerd, Denmark. pbra@novo.dk
Bjørn
S
S
Hastrup
S
S
Nielsen
P F
PF
Norris
K
K
Thim
L
L
Wiberg
F C
FC
Flodgaard
H
H
D000818:Animals; D023181:Antimicrobial Cationic Peptides; D016367:Baculoviridae; D001798:Blood Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology*; D002352:Carrier Proteins; D002460:Cell Line; D002634:Chemotaxis, Leukocyte / Q000187:drug effects*; D016207:Cytokines / Q000096:biosynthesis*; D004926:Escherichia coli; D006801:Humans; D007249:Inflammation; D008070:Lipopolysaccharides / Q000494:pharmacology*; D013058:Mass Spectrometry; D009000:Monocytes / Q000187:drug effects / Q000276:immunology / Q000502:physiology*; D009504:Neutrophils / Q000502:physiology*; D016133:Polymerase Chain Reaction; D011498:Protein Precursors / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000494:pharmacology; D018411:Spodoptera; D014162:Transfection
C062991:AZU1 protein, human; D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; D002352:Carrier Proteins; D016207:Cytokines; D008070:Lipopolysaccharides; D011498:Protein Precursors; D011994:Recombinant Proteins