TY - JOUR
T1 - Profiling the extended cleavage specificity of the house dust mite protease allergens der p 1, der p 3 and der p 6 for the prediction of new cell surface protein substrates
AU - Jacquet, Alain
AU - Campisi, Vincenzo
AU - Szpakowska, Martyna
AU - Dumez, Marie Eve
AU - Galleni, Moreno
AU - Chevigné, Andy
N1 - Funding Information:
This work was supported by the ?Fonds National de la Recherche? (FNRS), the ?Fonds de la Recherche Fondamentale et Collective? (2.4.511.06, 2.4.561.07, and 2.4.548.10), and Interuniversity Attraction Pole (IAP) P6/19, the Ministry of Research of Luxembourg, Luxembourg Institute of Health (LIH) grants 20160116 and 20170113 and the ?Fonds National de la Recherche? (FNR) Luxembourg, grants AFR-3004509. Vincenzo Campisi and Marie-Eve Dumez were supported by the Fonds pour la formation a la Recherche dans l?Industrie et dans l?Agriculture (FRIA) (Brussels, Belgium). Alain Jacquet is supported by National Research University Project, Office of Higher Education Commission (NRU59-003-HR) as well as by Chulalongkorn Academic Advancement (2nd Century Project-CUAASC). The authors would like to thank Julie Mathu, Nadia Beaupain and Manuel Counson for their technical assistance.
Publisher Copyright:
© 2017 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2017/7
Y1 - 2017/7
N2 - House dust mite (HDM) protease allergens, through cleavages of critical surface proteins, drastically influence the initiation of the Th2 type immune responses. However, few human protein substrates for HDM proteases have been identified so far, mainly by applying time-consuming target-specific individual studies. Therefore, the identification of substrate repertoires for HDM proteases would represent an unprecedented key step toward a better understanding of the mechanism ofHDMallergic response. In this study, phage display screenings using totally or partially randomized nonameric peptide substrate libraries were performed to characterize the extended substrate specificities (P5–P4ʹ) of the HDM proteases Der p 1, Der p 3 and Der p 6. The bioinformatics interface PoPS (Prediction of Protease Specificity) was then applied to define the proteolytic specificity profile of each protease and to predict new protein substrates within the human cell surface proteome, with a special focus on immune receptors. Specificity profiling showed that the nature of residues in P1 but also downstream the cleavage sites (Pʹ positions) are important for effective cleavages by all three HDM proteases. Strikingly, Der p 1 and Der p 3 display partially overlapping specificities. Analysis with PoPS interface predicted 50 new targets for the HDM proteases, including 21 cell surface receptors whose extracellular domains are potentially cleaved by Der p 1, Der p 3 and/or Der p 6. Twelve protein substrate candidates were confirmed by phage ELISA (enzyme linked immunosorbent assay). This extensive study of the natural protein substrate specificities of the HDM protease allergens unveils new cell surface target receptors for a better understanding on the role of these proteases in the HDM allergic response and paves the way for the design of specific protease inhibitors for future anti-allergic treatments.
AB - House dust mite (HDM) protease allergens, through cleavages of critical surface proteins, drastically influence the initiation of the Th2 type immune responses. However, few human protein substrates for HDM proteases have been identified so far, mainly by applying time-consuming target-specific individual studies. Therefore, the identification of substrate repertoires for HDM proteases would represent an unprecedented key step toward a better understanding of the mechanism ofHDMallergic response. In this study, phage display screenings using totally or partially randomized nonameric peptide substrate libraries were performed to characterize the extended substrate specificities (P5–P4ʹ) of the HDM proteases Der p 1, Der p 3 and Der p 6. The bioinformatics interface PoPS (Prediction of Protease Specificity) was then applied to define the proteolytic specificity profile of each protease and to predict new protein substrates within the human cell surface proteome, with a special focus on immune receptors. Specificity profiling showed that the nature of residues in P1 but also downstream the cleavage sites (Pʹ positions) are important for effective cleavages by all three HDM proteases. Strikingly, Der p 1 and Der p 3 display partially overlapping specificities. Analysis with PoPS interface predicted 50 new targets for the HDM proteases, including 21 cell surface receptors whose extracellular domains are potentially cleaved by Der p 1, Der p 3 and/or Der p 6. Twelve protein substrate candidates were confirmed by phage ELISA (enzyme linked immunosorbent assay). This extensive study of the natural protein substrate specificities of the HDM protease allergens unveils new cell surface target receptors for a better understanding on the role of these proteases in the HDM allergic response and paves the way for the design of specific protease inhibitors for future anti-allergic treatments.
KW - Allergen
KW - Cell surface proteome
KW - Dermatophagoides pteronyssinus
KW - House dust mite
KW - Phage display
KW - Phage substrate
KW - Protease
UR - http://www.scopus.com/inward/record.url?scp=85021419886&partnerID=8YFLogxK
U2 - 10.3390/ijms18071373
DO - 10.3390/ijms18071373
M3 - Article
C2 - 28654001
AN - SCOPUS:85021419886
SN - 1661-6596
VL - 18
JO - International Journal of Molecular Sciences
JF - International Journal of Molecular Sciences
IS - 7
M1 - 1373
ER -