Specific cleavage of the lung surfactant protein A by human cathepsin S may impair its antibacterial properties

Abstract

Human cysteine cathepsins (Cats) are implicated in lung injuries and tissue remodeling and have recently emerged as important players in pulmonary inflammations. The proteolytic activities of Cat B, L, K, S and H are dramatically increased in the sputum of patients with cystic fibrosis (CF), suggesting a possible involvement in the CF pathophysiology. We found that pulmonary surfactant protein A (SP-A) that participates to innate host defense is extensively degraded in CF expectorations. Breakdown of SP-A was markedly decreased in CF sputum by E-64 and Mu-Leu-Hph-VSPh, a Cat S inhibitor. Cat S cleaved efficiently and specifically SP-A within critical residues of the solvent-exposed loop of its carbohydrate recognition (C-type lectin) domain that allows binding to pathogens. Cat S decreased aggregation properties of SP-A (self-aggregation, aggregation of phospholipid vesicles and rough LPS). Moreover cleavage of SP-A by Cat S reduced binding to yeast mannan and impaired agglutination of Escherichia coli and Pseudomonas aeruginosa, a foremost detrimental pathogen colonizing the lungs of CF patients. Besides human neutrophil serine proteases and bacterial proteases, we propose that Cat S may participate in the pathophysiology of CF by weakening the antibacterial activity of SP-A. More broadly, present results provide further indication that Cat S, along with Cats B and L, could display immuno-modulatory functions by inactivating key proteins involved in the innate immunity defense.

Keywords: 1,2-dipalmitoyl-sn-glycero-3-phosphocholine; 1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol; 3-NO(2)-Tyr; 3-nitro-tyrosine; 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride; 7-amino-4-methyl coumarin; AEBSF; AMC; Abz; Antimicrobial protein; BALF; BCA; CA-074; CF; Cat; Collectin; Cysteine protease; Cystic fibrosis; DL-dithiothreitol; DPPC; DPPG; DTT; E-64; FRET; HNE; L-3-carboxy-trans-2.3-epoxypropionyl-leucylamido-(4-guanidino) butane; Lung; MMTs; Mu-Leu-Hph-VSPh; N-(l-3-trans-propylcarbamoyloxirane-2-carbonyl)-l-isoleucyl-l-proline; NSP; P. aeruginosa-negative; PMA; PMSF; Proteolysis; Ps+; Ps-; Pseudomonas aeruginosa-positive; RT; SP-A; TBS; Z; benzyloxycarbonyl; bicinchoninic acid; broncho-alveolar lavage fluid; cathepsin; cystic fibrosis; fluorescence resonance energy transfer; human neutrophil elastase; methylmethanethiosulfonate; morpholine urea-leucyl-homophenylalanyl-(vinylsulfonyl)benzene; neutrophil serine protease; ortho-aminobenzoic acid; phenylmethylsulfonyl fluoride; phorbol 12-myristate 13-acetate; room temperature; surfactant protein A; tris buffer saline.