The carbohydrate composition of pulmonary secretions

Potter, John; Matthews, Larry S.; Lemm, John; Spector, Samuel

doi:10.1016/s0022-3476(62)80280-7

Cited by 7 publications

(20 citation statements)

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“…Moreover, like bovine DNase I, rhDNase is inactivated by heat (100°C for 10 min), is specifically inhibited by actin (using a 1:1 molar ratio ofrhDNase and rabbit muscle actin), and shows optimal enzymatic activity at neutral pH (5.5-7.5) (39). Both the reported concentrations of Ca2+ and Mg2+ in sputum and the pH of sputum are sufficient to support the activity of rhDNase (40,41).…”

Section: Resultsmentioning

confidence: 90%

“…Experiments in the 1950s and 1960s revealed that DNA is present in very large amounts (3-14 mg/ml) in purulent, but not in uninfected, lung secretions (9)(10)(11). DNA, an extremely viscous polyanion, may contribute both to the increased viscosity of lung secretions and to the reduced effectiveness of aminoglycoside antibiotics (12)(13)(14). In fact, lung secretions incubated in vitro with partially purified bovine pancreatic DNase I (contaminated with proteases) show a large reduction in viscosity (15,16).…”

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confidence: 99%

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Recombinant human DNase I reduces the viscosity of cystic fibrosis sputum.

Shak¹,

Capon²,

Hellmiss³

et al. 1990

Proc. Natl. Acad. Sci. U.S.A.

650

399

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Respiratory distress and progressive lung destruction in cystic fibrosis can be attributed to bacterial persistence and the accumulation of viscous purulent secretions in the airways. More than 30 yr ago it was suggested that the large amounts of DNA in purulent secretions contribute to its viscosity and that bovine pancreatic DNase I could reduce the viscosity. To evaluate the potential clinical utility of recombinant human DNase I (rhDNase) in the treatment of cystic fibrosis, we have cloned, sequenced, and expressed rhDNase. Catalytic amounts of rhDNase greatly reduce the viscosity of purulent cystic fibrosis sputum, transforming it within minutes from a nonflowing viscous gel to a flowing liquid. The reduction in viscosity is associated with a decrease in size of DNA in the sputum. Inhalation of a rhDNase aerosol may be a simple direct approach that will help individuals with cystic fibrosis and other patients with pneumonia or bronchitis to clear their airways of purulent secretions.

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Section: Resultsmentioning

confidence: 90%

mentioning

confidence: 99%

Recombinant human DNase I reduces the viscosity of cystic fibrosis sputum.

Shak¹,

Capon²,

Hellmiss³

et al. 1990

Proc. Natl. Acad. Sci. U.S.A.

650

399

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“…During activation or phagocytosis, neutrophils release part of their cat G, elastase, and proteinase 3 content in airway secretions. In addition, when these short lived cells die in situ they release both their proteinase and DNA content (26). We have shown that DNA forms a tight complex with cat G, and so it renders this proteinase virtually resistant to inhibition by the fast acting serpins ␣ 1 PI and ACT.…”

Section: Discussionmentioning

confidence: 96%

DNA Strongly Impairs the Inhibition of Cathepsin G by α1-Antichymotrypsin and α1-Proteinase Inhibitor

Duranton

Boudier

Belorgey

et al. 2000

Journal of Biological Chemistry

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“…Cystic fibrosis sputum contains high amounts of DNA (4 mg/ml on the average) [14]. This DNA originates from neutrophils, forms a complex with mucus glycoproteins, is entrapped in the water-insoluble fraction of sputum and is responsible in part for the high viscosity of airways secretions which leads to airways obstruction [5,15]. The bars represent the standard errors calculated from repeated determinations.…”

Section: Discussionmentioning

confidence: 99%

“…Despite ~:his high load of inhibitor, bronchial secretions from patients ~vith cystic fibrosis contain significant amounts of active NE [4]. On the other hand, these secretions also contain large quantities ~f neutrophil-derived DNA [5]. This raises the question as to uhether DNA interferes with the inhibition of NE by MPI.…”

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confidence: 99%

DNA binds neutrophil elastase and mucus proteinase inhibitor and impairs their functional activity

Belorgey

Bieth

1995

FEBS Letters

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DNA binds neutrophil elastase and mucus proteinase inhibitor as evidenced by affinity chromatography on elastaseSepharose, inhibitor-Sepharose and DNA-cellulose. DNA is a potent hyperbolic inhibitor of elastase. The polynucleotide-en~yme complex is partially active on synthetic substrates and on elastin. DNA strongly increases kd~ and Ki for the inhibition of elastase by mucus proteinase inhibitor kass t E + I ~ EI, K i = kdi~Jka~s).kdiss ['he above effects are all salt-dependent. At physiological ionic strength, DNA is a potent inhibitor of the elastolytic activity of elastase and increases k~ and K~ for the elastase-mucus proteinase inhibitor interaction 160-fold and 100-fold, respectively.,~ey words': Elastase; Mucus proteinase inhibitor; Enzyme i:inetics; DNA; Cystic fibrosis L. IntroductionNeutrophil elastase (NE) is a 30-kDa cationic glycoprotein .vhose crystal structure is known. This serine proteinase cleaves t number of plasma and extracellular matrix proteins including ,.:lastin. It is stored in the azurophil granules of neutrophils from ,vhich it may be released following excessive phagocytosis or :ell death. Its extracellular proteolytic action is normally pre-,lented by protein proteinase inhibitors such as ~l-proteinase nhibitor, ~2-macroglobulin or mucus proteinase inhibitor MPI) (for a review see [1]). The latter is an 11.7-kDa unglyco~ylated cationic protein composed of two domains of similar ;ize and architecture ([2] and refs. therein).MPI is the most abundant NE inhibitor of airways secretions ~here it occurs in concentrations as high as 5/IM [3]. Despite ~:his high load of inhibitor, bronchial secretions from patients ~vith cystic fibrosis contain significant amounts of active NE [4]. On the other hand, these secretions also contain large quantities ~f neutrophil-derived DNA [5]. This raises the question as to uhether DNA interferes with the inhibition of NE by MPI. The ~resent investigation attempts to answer this question. Materials and methods MaterialsHuman NE was purified from purulent sputum as described in [6]. Recombinant MPI was obtained from Synergen, Boulder, CO through the courtesy of Dr. H.P. Schnebli, Ciba-Geigy, Basel, Switzerland. NE and MPI were active site titrated as indicated in [6]. Stock solutions of Suc-Ala3-pNA and MeOSuc-Ala2-Pro-Val-pNA (Bachem, Bubendorf, Switzerland) were made in N-methylpyrolidone and dimethylformamide, respectively. The final concentration of organic solvent in the reaction mixtures was 2% (v/v) throughout. Remazol Brilliant-blue elastin (RBB-elastin) was purchased from Elastin Products Company (Owenville, MO). Salmon sperm DNA and DNA~cellulose came from Sigma. NE or MPI were coupled to epoxy-activated Sepharose (Pharmacia) as described by the manufacturer. The three affinity supports (4 ml) were poured into HR 10/10 Pharmacia columns and equilibrated with 50 mM HEPES buffer, pH 7.4. Determination of k,,, and K,~The kinetic constants kca t and Km for the hydrolysis of MeOSuc-Ala2-Pro-Val-pNA by 30 nM free or DNA-bound NE were determine...

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The carbohydrate composition of pulmonary secretions

Cited by 7 publications

References 0 publications

Recombinant human DNase I reduces the viscosity of cystic fibrosis sputum.

Recombinant human DNase I reduces the viscosity of cystic fibrosis sputum.

DNA Strongly Impairs the Inhibition of Cathepsin G by α1-Antichymotrypsin and α1-Proteinase Inhibitor

DNA binds neutrophil elastase and mucus proteinase inhibitor and impairs their functional activity

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