Structural characterization of mouse neutrophil serine proteases and identification of their substrate specificities: relevance to mouse models of human inflammatory diseases.
It is widely accepted that neutrophil serine proteases (NSPs) play a critical role in neutrophil-associated lung inflammatory and tissue-destructive diseases. To investigate NSP pathogenic role(s), various mouse experimental models have been developed that mimic acutely or chronically injured human lungs. We and others are using mouse exposure to cigarette smoke as a model for chronic obstructive pulmonary disease with or without exacerbation. However, the relative contribution of NSPs to lung disease processes as well as their underlying mechanisms remains still poorly understood. And the lack of purified mouse NSPs and their specific substrates have hampered advances in these studies. In this work, we compared mouse and human NSPs and generated three-dimensional models of murine NSPs based on three-dimensional structures of their human homologs. Analyses of these models provided compelling evidence that peptide substrate specificities of human and mouse NSPs are different despite their conserved cleft and close structural resemblance. These studies allowed us to synthesize for the first time novel sensitive fluorescence resonance energy transfer substrates for individual mouse NSPs. Our findings and the newly identified substrates should better our understanding about the role of NSPs in the pathogenesis of cigarette-associated chronic obstructive pulmonary disease as well as other neutrophils-associated inflammatory diseases.
Kalupov
Timofey
T
INSERM U618, Protéases et Vectorisation Pulmonaires, IFR 135, Université François Rabelais de Tours, 37032 Tours, France.
Brillard-Bourdet
Michèle
M
Dadé
Sébastien
S
Serrano
Hélène
H
Wartelle
Julien
J
Guyot
Nicolas
N
Juliano
Luiz
L
Moreau
Thierry
T
Belaaouaj
Azzaq
A
Gauthier
Francis
F
D000818:Animals; D002403:Cathepsins / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D007249:Inflammation / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D008968:Molecular Conformation; D009504:Neutrophils / Q000201:enzymology* / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation; D057057:Serine Proteases / Q000737:chemistry* / Q000378:metabolism; D012907:Smoking / Q000009:adverse effects; D013045:Species Specificity; D013379:Substrate Specificity
D010455:Peptides; D002403:Cathepsins; D057057:Serine Proteases