title: ADP-ribosyltransferase-specific modification of human neutrophil peptide-1.
abstract: Epithelial cells lining human airways and cells recruited to airways participate in the innate immune response in part by releasing human neutrophil peptides (HNP). Arginine-specific ADP-ribosyltransferases (ART) on the surface of these cells can catalyze the transfer of ADP-ribose from NAD to proteins. We reported that ART1, a mammalian ADP-ribosyltransferase, present in epithelial cells lining the human airway, modified HNP-1, altering its function. ADP-ribosylated HNP-1 was identified in bronchoalveolar lavage fluid (BALF) from patients with asthma, idiopathic pulmonary fibrosis, or a history of smoking (and having two common polymorphic forms of ART1 that differ in activity), but not in normal volunteers or patients with lymphangioleiomyomatosis. Modified HNP-1 was not found in the sputum of cystic fibrosis patients or in leukocyte granules of normal volunteers. The finding of ADP-ribosyl-HNP-1 in BALF but not in leukocyte granules suggests that the modification occurred in the airway. Most of the HNP-1 in the BALF from individuals with a history of smoking was, in fact, mono- or di-ADP-ribosylated. ART1 synthesized in Escherichia coli, glycosylphosphatidylinositol-anchored ART1 released with phosphatidylinositol-specific phospholipase C from transfected NMU cells, or ART1 expressed endogenously on C2C12 myotubes modified arginine 14 on HNP-1 with a secondary site on arginine 24. ADP-ribosylation of HNP-1 by ART1 was substantially greater than that by ART3, ART4, ART5, Pseudomonas aeruginosa exoenzyme S, or cholera toxin A subunit. Mouse ART2, which is an NAD:arginine ADP-ribosyltransferase, was able to modify HNP-1, but to a lesser extent than ART1. Although HNP-1 was not modified to a significant degree by ART5, it inhibited ART5 as well as ART1 activities. Human beta-defensin-1 (HBD1) was a poor transferase substrate. Reduction of the cysteine-rich defensins enhanced their ability to serve as ADP-ribose acceptors. We conclude that ADP-ribosylation of HNP-1 appears to be primarily an activity of ART1 and occurs in inflammatory conditions and disease.
authors.affiliation: Pulmonary-Critical Care Medicine Branch, National Institutes of Health, Bethesda, Maryland 20892-1590.
authors.forename: Gregorino
authors.initials: G
authors.lastname: Paone
authors.affiliation: Pulmonary-Critical Care Medicine Branch, National Institutes of Health, Bethesda, Maryland 20892-1590.
authors.forename: Linda A
authors.initials: LA
authors.lastname: Stevens
authors.affiliation: Laboratory of Biochemistry, NHLBI, National Institutes of Health, Bethesda, Maryland 20892-1590.
authors.forename: Rodney L
authors.initials: RL
authors.lastname: Levine
authors.affiliation: Pulmonary-Critical Care Medicine Branch, National Institutes of Health, Bethesda, Maryland 20892-1590.
authors.forename: Christelle
authors.initials: C
authors.lastname: Bourgeois
authors.affiliation: Pulmonary-Critical Care Medicine Branch, National Institutes of Health, Bethesda, Maryland 20892-1590.
authors.forename: Wendy K
authors.initials: WK
authors.lastname: Steagall
authors.affiliation: Pulmonary-Critical Care Medicine Branch, National Institutes of Health, Bethesda, Maryland 20892-1590.
authors.forename: Bernadette R
authors.initials: BR
authors.lastname: Gochuico
authors.affiliation: Pulmonary-Critical Care Medicine Branch, National Institutes of Health, Bethesda, Maryland 20892-1590. Electronic address: mossj@nhlbi.nih.gov.
authors.forename: Joel
authors.initials: J
authors.lastname: Moss
mesh_terms: D036002:ADP Ribose Transferases / Q000737:chemistry*; D000818:Animals; D001249:Asthma / Q000378:metabolism; D001992:Bronchoalveolar Lavage Fluid; D003545:Cysteine / Q000737:chemistry; D003550:Cystic Fibrosis / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D058851:GPI-Linked Proteins; D006801:Humans; D007962:Leukocytes / Q000378:metabolism; D051379:Mice; D011658:Pulmonary Fibrosis; D012907:Smoking; D014162:Transfection; D023084:alpha-Defensins / Q000737:chemistry*
chemical_list: D058851:GPI-Linked Proteins; D023084:alpha-Defensins; C406413:human neutrophil peptide 1; D036002:ADP Ribose Transferases; C449301:ART1 protein, human; D003545:Cysteine