A dual-purpose synthetic colloidal platform for protease mapping: substrate profiling for Dengue and West Nile virus proteases.
In a proof of concept study, we created a small focused fluorescent hexapeptide library onto 14 multiplexed barcoded sets of silica particles to probe the substrate recognition specificity of West Nile and Dengue virus proteases. A flow cytometric analysis demonstrated that the optical signature of each bead population remained distinguishable throughout the solid-phase peptide synthesis and proteolytic assay. As expected, both proteases displayed a narrow specificity for lysine and arginine residues in the P(1) and P(2) substrate positions. This open-ended platform enables the fast and simultaneous identification of peptide substrates and is applicable to other proteases.
Marcon
Lionel
L
Nanotechnology and Biomaterials Centre, Level 5 East, Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Saint Lucia, QLD 4072, Australia. l.marcon@uq.edu.au
Kozak
Darby
D
Battersby
Bronwyn J
BJ
Chappell
Keith J
KJ
Fairlie
David P
DP
Young
Paul
P
Trau
Matt
M
D001120:Arginine / Q000378:metabolism; D003716:Dengue Virus / Q000201:enzymology*; D005434:Flow Cytometry; D008239:Lysine / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism*; D019151:Peptide Library; D013379:Substrate Specificity; D014764:Viral Proteins / Q000032:analysis / Q000378:metabolism*; D014902:West Nile virus / Q000201:enzymology*
D019151:Peptide Library; D014764:Viral Proteins; D001120:Arginine; D010447:Peptide Hydrolases; D008239:Lysine