Evaluation of the substrate specificity of human mast cell tryptase beta I and demonstration of its importance in bacterial infections of the lung.
Human pulmonary mast cells (MCs) express tryptases alpha and beta I, and both granule serine proteases are exocytosed during inflammatory events. Recombinant forms of these tryptases were generated for the first time to evaluate their substrate specificities at the biochemical level and then to address their physiologic roles in pulmonary inflammation. Analysis of a tryptase-specific, phage display peptide library revealed that tryptase beta I prefers to cleave peptides with 1 or more Pro residues flanked by 2 positively charged residues. Although recombinant tryptase beta I was unable to activate cultured cells that express different types of protease-activated receptors, the numbers of neutrophils increased >100-fold when enzymatically active tryptase beta I was instilled into the lungs of mice. In contrast, the numbers of lymphocytes and eosinophils in the airspaces did not change significantly. More important, the tryptase beta I-treated mice exhibited normal airway responsiveness. Neutrophils did not extravasate into the lungs of tryptase alpha-treated mice. Thus, this is the first study to demonstrate that the two nearly identical human MC tryptases are functionally distinct in vivo. When MC-deficient W/W(v) mice were given enzymatically active tryptase beta I or its inactive zymogen before pulmonary infection with Klebsiella pneumoniae, tryptase beta I-treated W/W(v) mice had fewer viable bacteria in their lungs relative to zymogen-treated W/W(v) mice. Because neutrophils are required to combat bacterial infections, human tryptase beta I plays a critical role in the antibacterial host defenses of the lung by recruiting neutrophils in a manner that does not alter airway reactivity.
Huang
C
C
Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts 02115, USA.
De Sanctis
G T
GT
O'Brien
P J
PJ
Mizgerd
J P
JP
Friend
D S
DS
Drazen
J M
JM
Brass
L F
LF
Stevens
R L
RL
D000595:Amino Acid Sequence; D000818:Animals; D006801:Humans; D007710:Klebsiella Infections / Q000201:enzymology / Q000473:pathology; D007711:Klebsiella pneumoniae / Q000201:enzymology; D008168:Lung / Q000201:enzymology* / Q000473:pathology; D008407:Mast Cells / Q000201:enzymology* / Q000473:pathology; D051379:Mice; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000473:pathology; D018410:Pneumonia, Bacterial / Q000201:enzymology* / Q000473:pathology; D012697:Serine Endopeptidases / Q000378:metabolism*; D013379:Substrate Specificity; D053802:Tryptases
C509190:Tpsb2 protein, mouse; D012697:Serine Endopeptidases; C509222:Tpsab1 protein, mouse; D053802:Tryptases