SP-A binds alpha1-antitrypsin in vitro and reduces the association rate constant for neutrophil elastase
© Gorrini et al. 2005
Received: 06 September 2005
Accepted: 13 December 2005
Published: 13 December 2005
α1-antitrypsin and surfactant protein-A (SP-A) are major lung defense proteins. With the hypothesis that SP-A could bind α1-antitrypsin, we designed a series of in vitro experiments aimed at investigating the nature and consequences of such an interaction.
Methods and results
At an α1-antitrypsin:SP-A molar ratio of 1:1, the interaction resulted in a calcium-dependent decrease of 84.6% in the association rate constant of α1-antitrypsin for neutrophil elastase. The findings were similar when SP-A was coupled with the Z variant of α1-antitrypsin. The carbohydrate recognition domain of SP-A appeared to be a major determinant of the interaction, by recognizing α1-antitrypsin carbohydrate chains. However, binding of SP-A carbohydrate chains to the α1-antitrypsin amino acid backbone and interaction between carbohydrates of both proteins are also possible. Gel filtration chromatography and turnover per inactivation experiments indicated that one part of SP-A binds several molar parts of α1-antitrypsin.
We conclude that the binding of SP-A to α1-antitrypsin results in a decrease of the inhibition of neutrophil elastase. This interaction could have potential implications in the physiologic regulation of α1-antitrypsin activity, in the pathogenesis of pulmonary emphysema, and in the defense against infectious agents.
Alpha1-antitrypsin (α1-AT) and surfactant protein-A (SP-A) are major defense glycoproteins in the alveolar spaces of human lungs. α1-AT, a 52,000 D glycoprotein, is secreted mostly by hepatocytes, and, to a lesser extent, by lung epithelial cells and phagocytes. α1-AT inhibits a variety of serine proteinases by its active site (Met358-Ser359), but its preferential target is human neutrophil elastase (HNE) as demonstrated by the high association rate constant (K ass) for this proteinase . In the lungs, α1-AT protects the connective tissue from HNE released by triggered neutrophils; as a result, subjects homozygous for the common deficiency variant Z α1-AT (associated with 15% of normal plasma α1-AT levels) develop pulmonary emphysema early in life, especially if they smoke .
SP-A, a member of the collectin (collagen-lectin) family , is one of the proteins of surfactant. Structurally, it comprises an N-terminal collagen-like domain connected by a neck to a C-terminal carbohydrate recognition domain (CRD) . Six trimers are linked by disulfide bridges in an octadecamer of 650,000 D, in a "flower bouquet" alignment pattern [4, 5]. A complex, predominantly triantennary, carbohydrate chain of ~4,000 D  is attached to the asparagine at position 187 of the CRD . SP-A is mainly present in the alveoli in association with phospholipids, only 1% being present in the free form [8, 9]. The primary function of surfactant is to reduce alveolar surface tension at end expiration. It is now however clear that SP-A, together with SP-D, another hydrophilic surfactant protein, plays a major role in the innate defenses of lung [5–10]. SP-A, in particular, is able to bind several micro-organisms and enhance their uptake by phagocytes, stimulate the production of free oxygen radicals, and induce phagocyte chemotaxis .
Most binding to micro-organisms, including influenza and herpes simplex viruses, Gram-positive and Gram-negative bacteria, mycobacteria, fungi, and Pneumocystis carinii, occurs via the CRD and is inhibited by sugars or calcium chelators .
Since some SP-A is present in the alveoli in the free form, it has a chance of coming into contact with α1-AT. We hypothesized that, in analogy with what happens with infectious agents, SP-A could bind to α1-AT, which carries 3 biantennary or triantennary asparagine-linked carbohydrate chains .
In this paper we provide in vitro evidence that the inhibitory activity of α1-AT towards HNE is significantly decreased in the presence of SP-A, probably as a consequence of SP-A binding to α1-AT. Such an interaction would represent a novel mechanism of regulating alveolar α1-AT. This could have relevance both for the pathogenesis of emphysema in patients with the Z α1-AT variant and for the lungs' defenses against infectious agents.
All reagents were of analytical grade, unless otherwise specified. The buffer used in all experiments was 0.2 M Na-K phosphate, with 0.5 M NaCl, 2 mM CaCl2, and 0.05% w/w Triton × 100, pH 8.0 (phosphate buffer), unless otherwise specified. Lipopolysaccharide (LPS) from E. coli serotype 026:B6 (Sigma) and methyl-α-D-mannopyranoside (MNOCH3) (Sigma) were dissolved in phosphate buffer. HNE and human α chymotrypsin (αChy) (ART, Athens, GA) were dissolved in 50 mM sodium acetate, 150 mM NaCl, pH 5.5 and diluted with phosphate buffer. N-glycocosidase F from Flavobacterium meningosepticum (PNGase F; EC 184.108.40.206) was purchased from Roche Diagnostics (Monza, Italy). Clostridium histolyticum collagenase type III (EC 3.4.24) came from Calbiochem (La Jolla, CA). The chromogenic substrates MeOSucAlaAlaProValNA (for HNE) and SucAlaAlaProPheNA (for αChy), and the irreversible inhibitors MeOSucAlaAlaProValCMK (for HNE) and TosPheCMK (for αChy) (all from Sigma) were dissolved in (CH3)2SO. Wild-type α1-antitryspin (M α1-AT) was either from ART or purified from human serum by covalent chromatography. Capillary isoelectric focusing (CIEF) with bare fused-silica capillaries filled with polyethylene oxide and carrier ampholyte solutions in the pH 3.5–5.0 range  was applied to confirm the presence of the common M α1-AT variant. Z α1-AT variant was purified by covalent chromatography from subjects identified within the Italian screening program for α1-AT deficiency . SP-A was isolated as described  from surfactant obtained from 3 patients affected by pulmonary alveolar proteinosis (PAP), subjected to therapeutic whole lung lavage  and from adult New Zealand rabbits. To isolate surfactant, the bronchoalveolar lavage fluid was filtered through gauze and centrifuged at 150 g for 10 minutes. The supernatant was centrifuged for 30 minutes at 80,000 × g and the resulting pellet was suspended in 10 mM Tris-HCl pH 7.4, 145 mM NaCl, 1.25 mM CaCl2, 1 mM MgCl2, 2.2 M sucrose (solution A), overlaid with 10 mM Tris-HCl pH 7.4, 145 mM NaCl, 1.25 mM CaCl2, 1 mM MgCl2, 2 M sucrose (solution B) and ultracentrifuged overnight at 85,000 × g in a Ti 60 rotor (Beckman). The floating material was dispersed in water and centrifuged for 30 minutes at 100,000 × g and the pellet recovered was stored at -70°C (purified surfactant). To obtain SP-A, surfactant was injected into a 50-fold excess by volume of 1-butanol and stirred at room temperature for 30 minutes. After centrifugation, the pellet was suspended in 1-butanol and re-centrifuged at 4,000 × g for 1 hour at room temperature. The final precipitate was dried under nitrogen and then resuspended in 5 mM Tris-HCl, 145 mM NaCl, 20 mM octyl β-D-glucopyranoside, pH 7.4 (solution C). After centrifugation at 100,000 × g for 1 hour, the pellet was resuspended in 5 mM Tris-HCl pH 7.4 (solution D) and dialyzed against solution D for 48 hours with at least six changes. The final solution was centrifuged at 100,000 × g for 1 hour and the resulting supernatant, containing purified SP-A, was stored. Endotoxin-free SP-A was obtained by treatment with polymyxin-B (Sigma). Small aliquots of SP-A in solution D were incubated in a 1:1 ratio for 6 hours at 4°C with polymyxin-agarose previously equilibrated with 5 mM Tris-HCl, 100 mM octyl β-D-glucopyranoside and 2 mM EDTA, pH 7.4. Polymyxin-agarose was removed by centrifugation at 14,000 × g for 15 minutes, and the supernatant was then dialyzed against 5 mM Tris-HCl pH 7.4 for 48 hours with at least six changes and lyophilized [17, 18]. For some experiments polymyxin-treated SP-A was further purified by D-mannose sepharose 4B chromatography. SP-A was added to a small column containing D-mannose sepharose 4B (Pharmacia) previously equilibrated with 5 mM HEPES, 0.4% Triton × 100 and 1.5 mM CaCl2, pH 7.2 (solution E), and the column was washed extensively with solution E. SP-A was finally eluted with 5 mM HEPES, 0.4% Triton × 100 and 2.5 mM EDTA, pH 7.2 (solution F).
Modification of the native proteins
Identification of the SP-A/ α1-AT complex
1) Gel filtration HPLC
A mixture of SP-A (1.62 mg/ml) and α1-AT (1 mg/ml) in a 1:50 molar ratio was incubated for 24 hrs at 37°C in phosphate buffer. The SP-A/α1-AT mixture and single proteins were loaded in a Jasco PU 980 HPLC system (Japan Spectroscopic, Tokyo, Japan) equipped with two Biosep-SEC-S 4000 columns (300 × 7.80 mm each, Phenomenex, Torrence, CA, USA) connected in series. Samples were eluted with 100 mM Na2HPO4, 2 mM CaCl2, pH 6.8 at a flow rate of 0.3 ml/ min, and monitored at 220 nm. The excluded (V0 = 12.43 ml) and total (Vt = 24.82 ml) volumes were determined using dextran and creatinine, respectively; a calibration curve was obtained by running through the column a set of standard proteins: α2-macroglobulin (725 kD), aldolase (158 kD), bovine serum albumin (67 kD), chymotrypsinogen (25 kD), and cytochrome C (12.5 kD). The results were reported as mean ± SD of three separate experiments. 2
2) Qualitative immunodetection by ELISA
250 ng of standard α1-AT, purified SP-A, and SP-A/α1-AT complex collected from the Size Exclusion Chromatography experiments, were immobilized in 50 mM Na2CO3, pH 9.5 overnight at 4°C in a polypropylene plate (Corning, New York, USA). Plates were then brought at room temperature, washed with 150 mM NaCl, 0.1% Tween 20 (ELISA buffer), blocked for 1 h with 50 mM Na2CO3, 2% BSA pH 9.5, incubated for 2 hrs in the presence of primary antibodies diluted 1:500 (goat anti-human α1-AT and rabbit anti-human SP-A; ICN, Aurora, OH, USA), washed and finally reacted for 2 hrs with the appropriate biotinylated secondary antibodies diluted 1:5000 (Chemicon, Temecula, CA, USA). After washing, 100 μL of avidin diluted 1:2000 were added, and samples were incubated for 30 min. Color development was achieved by incubating the samples with 1,2-phenylenediamine dihydrochloride (Dako, Bucks, UK). The reaction was stopped by addition of 100 μl of 0.5 M H2SO4 and OD was read at 490 nm with a Bio-Rad 680 Microplate Reader (Bio-Rad Laboratories, CA, USA).
Rate constants were derived by competition experiments of HNE and αChy. Kinetic parameters were determined as described [20, 21]. The active sites of HNE and αChy were titrated using a procedure based on the measurement of pNa released after enzymatic cleavage of MeOSucAlaAlaProValNA and SucAlaAlaProPheNA, respectively, at 37°C . Product formation was monitored spectrophotometrically at a wavelength of 405 nm using a Bio Rad Microplate Reader model 3550. To titrate the different forms of α1-ATs (α1-AT, deglycosylated α1-AT and Z α1-AT), 7.5 nM HNE was incubated for 15 min at 37°C with 0–100 nM inhibitor, in the presence of 2 mM MeOSucAlaAlaProValNA. All following kinetic experiments were derived from α1-ATs and SP-A/α1-ATs complexes (obtained by incubation of α1-ATs, from 0 to 25 nM, with SP-A 15, 7.5, 1.5, 0.15 mM for 15 min at 37°C). See additional file 1 for more details.
Association rate constant (K ass M-1sec-1) for inhibition of HNE by α1-AT with SP-A. SP-A employed was both from humans affected by PAP or from rabbit, polymyxin treated and polymyxin-mannose treated. Data are means ± SD of three different experiments.
Kass, (M-1sec-1) means ± SD
3.40 ± 0.0079 × 107
3.40 ± 0.0079 × 107
3.40 ± 0.0079 × 107
3.40 ± 0.0079 × 107
1.84 ± 0.0577 × 107
1.84 ± 0.0580 × 107
1.86 ± 0.0565 × 107
1.82 ± 0.0585 × 107
1.70 ± 0.0623 × 107
1.70 ± 0.0631 × 107
1.72 ± 0.0618 × 107
1.68 ± 0.0620 × 107
5.20 ± 0.0483 × 106
5.22 ± 0.0480 × 106
5.00 ± 0.0478 × 106
5.40 ± 0.0490 × 106
4.30 ± 0.0513 × 106
4.30 ± 0.0520 × 106
4.30 ± 0.0520 × 106
4.40 ± 0.0498 × 106
Association rate constant for inhibition of HNE by α1-AT and Z α1-AT with SP-A. Data are means ± SD of experiments performed in triplicate with 3 different batches of human SP-A and1batch of rabbit SP-A.
decrease in K ass, n-fold
decrease in K ass, n-fold
Z α1-AT nM
3.40 ± 0.0079 × 107
9.80 ± 0.0032 × 106
1.84 ± 0.0577 × 107
5.20 ± 0.0314 × 106
1.70 ± 0.0623 × 107
4.70 ± 0.0268 × 106
5.20 ± 0.0483 × 106
4.30 ± 0.0240 × 106
4.30 ± 0.0513 × 106
3.80 ± 0.0221 × 106
To exclude that the results were due to non-specific binding, we incubated 7.5 nM HNE with 0–100 nM α1-AT for 15 min at 37°C in microtiter plates or in glass tubes and then measured the residual HNE activity with 2 mM MeOSucAlaAlaProValNA, finding no difference between plastics and glass. Furthermore, to exclude binding of SP-A to plastics we incubated 15 nM SP-A with I125α1-AT (from 0 to 100 nM) at 37°C. The number of Cpm of the samples with SP-A were the same of wells without proteins. We concluded that our data were compatible with binding of α1-AT to SP-A.
Deglycosylated α1-AT retains its ability to inhibit HNE (K ass 3.38 × 107 M-1sec-1). We did, however, find that the inhibitory activity of α1-AT is greatly decreased in the presence of SP-A (K ass 1.1 × 107 M-1sec-1, Figure 4), indicating that binding of SP-A to the carbohydrate moiety of α1-AT is not the only mechanism involved. The turnover number of the SP-A/deglycosylated α1-AT is 12, half that displayed by native α1-AT (Figure 4, 5). To explore other mechanisms of binding between SP-A and α1-AT, we incubated boiled SP-A and α1-AT. We found that boiled SP-A/native α1-AT displayed the same K ass and the same turnover number as native SP-A/deglycosylated α1-AT (Figures 4, 5). We postulated that SP-A carbohydrate chains could bind α1-AT, possibly through the amino acid backbone. In fact, carbohydrate chains isolated from SP-A mixed with deglycosylated α1-AT resulted in the same K ass and turnover number as those of native SP-A/deglycosylated α1-AT (Figures 4, 5). Besides these mechanisms of binding of SP-A to α1-AT, a third mechanism, i.e. a carbohydrate/carbohydrate interaction, probably exists since boiled SP-A and native α1-AT displayed a K ass of 1.9 × 107 M-1sec-1 and ~6 turnovers (Figure 4, 5).
Finally, we studied the binding of deglycosylated SP-A to α1-AT. The K ass of native α1-AT mixed with deglycosylated SP-A was 1.2 × 107 M-1sec-1 and the turnover number 18 (Figure 4, 5). Absence of SP-A/α1-AT binding, i.e. K ass 3.4 × 107 M-1sec-1, and a turnover number of 1, was achieved by two combinations: 1) SP-A deglycosylated and boiled with native α1-AT, and 2) both proteins deglycosylated (Figures 4, 5). In the former case, absence of SP-A carbohydrates and denaturation of CRDs hindered any possible binding of SP-A to native α1-AT. In the latter case, the binding was precluded by the absence of carbohydrates on both proteins, in spite of the presence of intact CRDs in the SP-A.
The present data provide evidence for an in vitro interaction between SP-A and α1-AT. These glycoproteins belong to two systems of the lung that are supposed to act independently: the surfactant system and the proteinase/proteinase inhibitor system. Nevertheless, evidence for possible links between the two systems does exist. As an example, it has been shown that SP-A may be digested by elastolytic enzymes [28, 29], and that inhalation of α1-AT in patients with cystic fibrosis may result in an increase of SP-A levels in bronchoalveolar lavage fluid (BALf) . In addition, SP-D induces the production of matrix metalloproteinases by human alveolar macrophages , whereas the cysteine proteinase cathepsin H is involved in the first N-terminal processing step of SP-C . The two systems may therefore interact in the lungs, both in physiologic and in pathologic pathways. The concentration of SP-A in the BALf of normal subjects is estimated to be ~277 nM . Since approximately 1% of total SP-A is present in the free form [8, 9], its concentration in BALf would be ~2.8 nM. Given that the concentration of α1-AT is ~5 μM , we reasoned that the two glycoproteins have a good chance of coming into contact during their life cycle.
Indeed our in vitro experiments indicate that the interaction between SP-A and α1-AT results in binding between them. This binding, which is calcium-dependent, appears to be complex since it could involve binding between the CRD of SP-A and carbohydrates on α1-AT, binding between SP-A carbohydrates and the protein backbone of α1-AT, and binding between the carbohydrate chains of both proteins.
Turnover per inactivation suggests that one part of SP-A binds 23 molar parts of α1-AT. Nevertheless, SP-A binds 11 molar parts of deglycosylated, fully active α1-AT (Figure 4, 5), thus suggesting a possible binding of SP-A carbohydrate chains to the amino acid backbone of α1-AT. Asn, to which carbohydrates of the native glycoprotein are linked , is a likely candidate. This hypothesis was confirmed by the results obtained with boiled SP-A and with isolated SP-A carbohydrate chains (Figure 4, 5). In support of this hypothesis, it has been reported that the binding of SP-A to influenza virus , herpes virus type 1 infected cells , and M. tuberculosis , involves N-linked carbohydrate chains on SP-A. Interestingly, there may be multiple binding sites on individual micro-organisms .
Our experiments also suggest a possible carbohydrate/carbohydrate interaction between SP-A and α1-AT. Such a type of linkage has been shown to operate in the calcium-mediated homotypic interaction between two Lewis (Lex) determinants (Galβ1→4[Fucα1→3]GlcNAc) involved in cell adhesion during murine embryogenesis . Interestingly Lex-Lex interactions appear to be calcium-dependent , by involving van der Waal forces. The fact that ultra-weak interactions are involved explains why this aspect is often underestimated [39–41].
It is difficult to postulate whether the three proposed mechanisms of binding take place simultaneously between native proteins. It may be that the CRD plays the main role and that the other two mechanisms are less important or take place only as artificial mechanisms once the proteins have been manipulated.
The binding with SP-A results in a decrease in the inhibition of HNE by α1-AT. There are several known mechanisms that could explain the inactivation of α1-AT. Beside the physiologic irreversible suicide substrate mechanism by which α1-AT inhibits HNE , α1-AT may also be inactivated by oxidation of methionine residue(s) located at or near the active site [22, 23]. Another mechanism of α1-AT inactivation is proteolytic degradation at or near the active site by a number of host and non-host, mostly microbial, proteinases . Whether these mechanisms may act in vivo, thereby contributing to the imbalance between proteinases and inhibitors in the pathogenesis and progression of pulmonary emphysema, is still a debated issue.
With respect to the inhibitory activity of α1-AT, that of Z α1-AT is further impaired by this latter's enhanced tendency to undergo spontaneous polymerization . This phenomenon, also known as loop-sheet polymerization, likely accounts for why Z α1-AT is less efficient at inhibiting HNE, and has been demonstrated to be present in vivo, since Z α1-AT polymers have been detected in the BALf of Z α1-AT subjects with emphysema . We found that SP-A binds Z α1-AT and that the binding further reduces the K ass, which is already impaired with respect to that of α1-AT. Were this binding to happen in vivo, it would further decrease the antiproteinase activity of Z α1-AT.
In spite of the detailed dissection of the binding mechanism of SP-A to α1-AT in vitro, an obvious limitation of the present paper is the lack of specific studies investigating a possible interaction between SP-A and α1-AT in vivo. Nevertheless, some indirect evidence suggesting that such an interaction might take place is available, although it is not possible to address a plausible expectation of physiologic or pathophysiologic relevance of these findings. For example, a recent report has shown that in human sputum supramolecular complexes with heparan sulfate/Syndecan-1 and proteinase and inhibitors are present . These complexes contain the proteinase inhibitors SLPI and α1-AT, NE as well, whose proteolytic activity is however not decreased . The large MW of SP-A makes difficult to highlight the occurrence of such supramolecular complexes including α1-AT by standard techniques . Nevertheless, a report focusing on two-dimensional electrophoretic characteristics of BALf proteins in subjects affected by interstitial lung diseases  has intriguingly shown that some α1-AT fragments were superimposed on spots of SP-A, in its upper, acidic position. These findings, confirmed by mass spectrometric MALDI-TOF analysis, would suggest a possible SP-A/α1-AT interaction taking place in vivo.
We have shown that SP-A binds α1-AT, and that this binding results in a significant decrease in the association rate constant of α1-AT for HNE. The mechanism of the binding seems to be predominantly mediated by the SP-A CRDs, as indicated by the calcium dependence and by the turnovers for inactivation, but other mechanisms may be involved, such as an interaction between SP-A carbohydrates and the α1-AT amino acid backbone or between carbohydrate chains of both glycoproteins. The presence of these complex binding mechanisms would exclude the hypothesis that the α1-AT inhibition occurred simply due to steric inhibition of the large SP-A molecule, but it would rather suggest a programmed, coordinated mechanism.
The in vitro interaction described here, if present in vivo, would be a novel mechanism of impairment of α1-AT inhibitory activity. It might represent a physiologic mechanism of regulating α1-AT activity, especially in acute conditions (for example during defense against infections agents) , in which an excess of α1-AT would interfere with the physiologic role of proteinases. α1-AT is indeed a highly specialised proteinase inhibitor , but the presence in nature of several, robust mechanisms of α1-AT downregulation (i.e. inherited deficiency, susceptibility to oxidative stress and proteolysis, polymerization) would imply the occurrence of intrinsic risks related to the overexpression of a nearly perfect and immortal inhibitor. Therefore, the formation of supramolecular complexes SP-A/α1-AT might be a sort of reserve mechanism, taking place in case of need.
On the other hand, the interaction with SP-A would be of particular relevance in the pathogenesis of pulmonary emphysema associated with α1-AT deficiency, since it would contribute significantly to the complex mechanisms of imbalance between Z α1-AT and HNE in the lungs. Obviously, all these speculations need further investigations, first of all to understand whether or not SP-A/α1-AT binding is a relevant down-regulatory mechanism of α1-AT inhibitory activity in vivo.
carbohydrate recognition domain
human neutrophil elastase
- MNOCH3 :
We thank Dr. I. Ferrarotti for her contribution to this research, and Dr R. Stenner for editing the manuscript. This work was in part supported by the IRCCS Policlinico San Matteo Ricerca Corrente and by the Fondazione Cariplo
- Travis J, Salvesen GS: Human plasma proteinase inhibitors. Annu Rev Biochem 1983, 52:655–709.View ArticlePubMedGoogle Scholar
- Carrell RW, Lomas DA: Alpha 1 -antitrypsin deficiency. A model for conformational diseases. N Engl J Med 2002, 346:45–53.View ArticlePubMedGoogle Scholar
- Sastry K, Ezekowitz RA: Collectins: pattern recognition molecules involved in the first line of host defense. Curr Opin Immunol 1993, 5:59–63.View ArticlePubMedGoogle Scholar
- Haagsman HP, White RT, Schilling J, Lau K, Benson BJ, Golden J, Hawgood S, Clements JA: Studies of the structure of lung surfactant protein SP-A. Am J Physiol 1989, 257:L421-L429.PubMedGoogle Scholar
- McCormack FX, Whitsett JA: The pulmonary collectins, SP-A and SP-D, orchestrate innate immunity in the lung. J Clin Invest 2002, 109:707–712.View ArticlePubMedPubMed CentralGoogle Scholar
- Bhattacharyya SN, Lynn WS: Structural studies and oligosaccharides of glycoprotein isolated from alveoli of patients with alveolar proteinosis. J Biol Chem 1977, 252:1172–1180.PubMedGoogle Scholar
- Munakata H, Nimberg RB, Snider GL, Robins AG, Van Halbeek H, Vliegenthart JF, Schmid K: The structure of the carbohydrate units of the 36K glycoprotein derived from the lavage of a patient with alveolar proteinosis by high-resolution 1 H-NMR spectroscope. Biochem Biophys Res Commun 1982, 108:1401–1405.View ArticlePubMedGoogle Scholar
- Baritussio A, Alberti A, Quaglino D, Pettenazzo A, Dal Zoppo D, Sartori L, Pasquali-Ronchetti I: SP-A, SP-B, and SP-C in surfactant subtypes around birth: reexamination of alveolar life cycle of surfactant. Am J Physiol 1994, 266:L436-L447.PubMedGoogle Scholar
- Savov J, Wright JR, Young SL: Incorporation of biotinylated SP-A into rat lung surfactant layer, type II cells, and Clara cells. Am J Physiol 2000, 279:L118-L126.Google Scholar
- Wright JR: Immunomodulatory functions of surfactant. Physiol Rev 1997, 77:931–962.PubMedGoogle Scholar
- Korfhagen TR: Surfactant protein A (SP-A)-mediated bacterial clearance. SP-A and cystic fibrosis. Am J Respir Cell Mol Biol 2001, 25:668–672.View ArticlePubMedGoogle Scholar
- Mason RJ, Greene K, Voelker DR: Surfactant protein A and surfactent protein D in health and disease. Am J Physiol 1998, 275:L1-L13.PubMedGoogle Scholar
- Mega T, Lujan E, Yoshida A: Studies on the oligosaccharide chains of human α1-proteinase inhibitor. I. Isolation of glycopeptides. J Biol Chem 1980, 255:4053–4056.PubMedGoogle Scholar
- Lupi A, Viglio S, Luisetti M, Gorrini M, Coni P, Faa G, Cetta G, Iadarola P: α1-antitrypsin in serum determined by capillary isoelectric focusing. Electrophoresis 2000, 21:3318–3326.View ArticlePubMedGoogle Scholar
- Ferrarotti I, Baccheschi J, Zorzetto M, Tinelli C, Corda L, Balbi B, Campo I, Pozzi E, Faa G, Coni P, Massi G, Stell G, Luisetti M: Prevalence and phenotype of subjects carrying rare variants in the Italian Registry for alpha1-antitrypsin deficiency. J Med Genet 2005, 42:282–287.View ArticlePubMedPubMed CentralGoogle Scholar
- Howgood S, Benson BJ, Shilling J, Damm D, Clements JA, White RT: Nucleotide and amino acid sequence of pulmonary surfactant protein SP18 and evidence for co-operation between SP18 and SP28–36 in surfactant lipid adsorption. Pro Natl Acad Sci USA 1987, 84:66–70.View ArticleGoogle Scholar
- Alberti A, Luisetti M, Braschi A, Rodi G, Iotti G, Sella D, Poletti V, Benori V, Baritussio A: Broncho-alveolar lavage fluid composition in alveolar proteinosis. Early changes after therapeutic lavage. Am J Respir Crit Care Med 1996, 154:817–820.View ArticlePubMedGoogle Scholar
- Wright JR, Zlogar DF, Taylor JC, Zlogar TM, Restepo CI: Effect of endotoxin on surfactant protein A and D stimulation of NO production by alveolar macrophages. Am J Physiol 1999, 276:L650-L658.PubMedGoogle Scholar
- Meloni F, Alberti A, Bulgheroni A, Lupi A, Paschetto E, Marone Bianco A, Rodi G, Fietta A, Luisetti M, Baritussio A: Surfactant apoprotein A modulates interleukin-8 and monocyte chemotactic peptide-1 production. Eur Respir J 2002, 19:1128–1135.View ArticlePubMedGoogle Scholar
- Vincent J-P, Lazdunski M: Trypsin-pancreatic trypsin inhibitor associations. Dynamics of the interaction and role of disulfide bridges. Biochemistry 1972, 11:2967–2977.View ArticlePubMedGoogle Scholar
- Beatty K, Bieth J, Travis J: Kinetics of association of serine proteinases with native and oxidized α-1-proteinase inhibitor and α-1-antichymotrypsin. J Biol Chem 1980, 255:3931–3934.PubMedGoogle Scholar
- Gorrini M, Lupi A, Viglio S, Pamparana F, Cetta G, Iadarola P, Powers JC, Luisetti M: Inhibition of human neutrophil elastase by erythromycin and flurythromycin, two macrolide antibiotics. Am J Respir Cell Mol Biol 2001, 25:492–499.View ArticlePubMedGoogle Scholar
- Creuwels LAJM, van Golde LMG, Haagsman H: The pulmonary surfactant system: biochemical and clinical aspects. Lung 1997, 175:1–39.View ArticlePubMedGoogle Scholar
- Ogushi F, Fells GA, Hubbard RC, Straus SD, Crystal RG: Z-type α1-antitrypsin is less competent than M1-type α1-antitrypsin as an inhibitor of neutrophil elastase. J Clin Invest 1987, 80:1366–1374.View ArticlePubMedPubMed CentralGoogle Scholar
- Haagsman HP, Sargeant T, Hauschka VH, Benson BJ, Hawgood S: Binding of calcium to SP-A, a surfactant-associated protein. Biochemistry 1990, 29:8894–8900.View ArticlePubMedGoogle Scholar
- Haurum JS, Thiel S, Haagsman HP, Laursen SB, Larsen B, Jensenius JC: Studies on the carbohydrate-binding characteristics of human pulmonary surfactant-associated protein A and comparison with two other collectins: mannan-binding protein and conglutinin. Biochem J 1993, 293:873–87.View ArticlePubMedPubMed CentralGoogle Scholar
- Khubchandani KR, Oberley RE, Snyder JM: Effects of surfactant protein A and NaCl concentration on the uptake of Pseudomonas aeruginosa by THP-1 cells. Am J Respir Cell Mol Biol 2001, 25:699–706.View ArticlePubMedGoogle Scholar
- Rubio F, Cooley J, Accurso FJ, Remold-O'Donnell E: Linkage of neutrophil serine proteases and decreased surfactant protein-A (SP-A) levels in inflammatory lung disease. Thorax 2004, 59:318–323.View ArticlePubMedPubMed CentralGoogle Scholar
- Beatty AL, Malloy JL, Wright JR: Pseudomonas aeruginosa degrades pulmonary surfactant and increases conversion in vitro . Am J Respir Cell Mol Biol 2005, 32:128–134.View ArticlePubMedGoogle Scholar
- Griese M, von Bredow C, Birrer P: Reduced proteolysis of surfactant protein A and changes of the bronchoalveolar lavage fluid proteome by inhaled alpha 1-protease inhibitor in cystic fibrosis. Electrophoresis 2001, 22:165–171.View ArticlePubMedGoogle Scholar
- Crippes Trask B, Malone MJ, Lum EH, Welgus HG, Crouch EC, Shapiro SD: Induction of macrophage matrix metalloproteinase biosynthesis by surfactant protein D. J Biol Chem 2001, 276:37846–37852.Google Scholar
- Brasch F, ten Brinke A, Johnen G, Ochs M, Kapp N, Müller KM, Beers MF, Fehrenbach H, Richter J, Batenburg JJ, Bühling F: Involvement of cathepsin H in the processing of the hydrophobic surfactant-associated protein C in type II pneumocytes. Am J Respir Cell Mol Biol 2002, 26:659–670.View ArticlePubMedGoogle Scholar
- Van de Graaf EA, Jansen HM, Lutter R, Alberts C, Kobsen J, de Vries IJ, Out TA: Surfactant protein A in bronchoalveolar lavage fluid. J Lab Clin Med 1992, 120:252–263.PubMedGoogle Scholar
- Rennard SI, Ghafouri M, Thompson AB, Linder J, Vaughan W, Jones K, Ertl RF, Christensen K, Prine A, Stahl MG, et al.: Fractional processing of sequential bronchoalveolar lavage to separate bronchial and alveolar samples. Am Rev Respir Dis 1990, 141:208–217.View ArticlePubMedGoogle Scholar
- Sprio RG: Protein glycosylation: nature, distribution, enzymatic formation, and disease implications of glycopeptide bonds. Glycobiology 2000, 12:43R-56R.View ArticleGoogle Scholar
- Benne CA, Kraaijeveld CA, Van Strijp JAG, Brouwer E, Harmsen M, Verhoef J, van Golde LMG, van Iwaarden JF: Interactions of surfactant protein A with influenza A viruses: binding and neutralization. J Infect Dis 1995, 171:335–341.View ArticlePubMedGoogle Scholar
- Van Iwaarden JF, Van Strijp JAG, Visser H, Haagsman HP, Verhoef J, van Golde LMG: Binding of surfactant protein A (SP-A) to herpes simplex virus type 1-infected cells is mediated by the carbohydrate moiety of SP-A. J Biol Chem 1992, 267:25039–25043.PubMedGoogle Scholar
- Gaynor CD, McCormack FX, Voelker DR, McGowan SE, Schlesinger LS: Pulmonary surfactant protein A mediates enhanced phagocytosis of Mycobacterium tuberculosis by a direct interaction with human macrophages. J Immunol 1995, 155:5343–5351.PubMedGoogle Scholar
- Pincet F, Le Bouar T, Zhang Y, Esnault J, Mallet J-M, Perez E, Sinaÿ P: Ultraweak sugar-sugar interactions for transient cell adhesion. Biophys J 2001, 80:1354–1358.View ArticlePubMedPubMed CentralGoogle Scholar
- Henry B, Desvaux H, Pritstchepa M, Berthault P, Zhang Y, Mallet J-M, Esnault J, Sinaÿ P: NMR study of a Lewis x pentasaccharide derivative: solution structure and interaction with cations. Carbohydr Res 1999, 315:48–62.View ArticlePubMedGoogle Scholar
- Sears P, Wong C-H: Intervention of carbohydrate recognition by proteins and nucleic acids. Proc Natl Acad Sci USA 1996, 93:12086–12093.View ArticlePubMedPubMed CentralGoogle Scholar
- Silverman GA, Bird PI, Carrell RW, Church FC, Coughlin PB, Gettings PGW, Irving JA, Lomas DA, Luke CJ, Moyer RW, Pemberton PA, Remold-O'Donnell E, Salvesen GS, Travis J, Whisstock JC: The serpins are an expanding superfamily of structurally similar but functionally diverse proteins. J Biol Chem 2001, 276:33293–33296.View ArticlePubMedGoogle Scholar
- Taggart C, Cervantes-Laurean D, Kim G, McElvaney NG, Wehr N, Moss J, Levine RL: Oxidation of either methionine 351 or methionine 358 in α1-antitrypsin causes loss of anti-neutrophil elastase activity. J Biol Chem 2000, 275:27258–27265.PubMedGoogle Scholar
- Travis J, Shieh B-H, Potempa J: The functional role af acute phase plasma proteinase inhibitors. Tokai J Exp Clin Med 1988, 13:313–320.PubMedGoogle Scholar
- Elliott P, Bilton D, Lomas DA: Lung polymers in Z α 1 -antitrypsin related emphysema. Am J Respir Cell Mol Biol 1998, 18:670–674.View ArticlePubMedGoogle Scholar
- Chan SCH, Shum DKY, Ip MSM: Sputum sol neutrophil elastase activity in bronchiectasis. Differential modulation by Syndecan-1. Am J Respir Crit Care Med 2003, 168:192–198.View ArticlePubMedGoogle Scholar
- Madsen J, Kliem A, Nielsen O, Koch C, Steinhilber W, Holmskov U: Expression and localization of lung surfactant protein A in human tissues. Am J Respir Cell Mol Biol 2003, 29:591–597.View ArticlePubMedGoogle Scholar
- Magi B, Bini L, Perari MG, Fossi A, Sanchez JC, Hochstrasser D, Paesano S, Raggiaschi R, Santucci A, Pallini V, Rottoli P: Bronchoalveolar lavage fluid protein composition in patients with sarcoidosis and idiopathic pulmonary fibrosis: a two-dimensional electrophoretic study. Electrophoresis 2002, 23:3434–3444.View ArticlePubMedGoogle Scholar
- Matthay MA, Zimmerman GA: Centennial Review. Acute lung injury and the acute respiratory distress syndrome. Four decades of Inquiry into pathogenesis and rational management. Am J Respir Cell Mol Biol 2005, 33:319–327.View ArticlePubMedPubMed CentralGoogle Scholar
- Otlewski J, Jelen F, Zakreweska M, Oleksy A: The many faces of protease-protein inhibitor interaction. EMBO J 2005, 24:1303–1310.View ArticlePubMedPubMed CentralGoogle Scholar
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