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Designing Fluorescent Peptide Sensors with Dual Specificity for the Detection of HIV‑1 Protease
journal contribution
posted on 2015-10-27, 00:00 authored by Karla-Luise Herpoldt, Arbel Artzy-Schnirman, Andrew J. Christofferson, Adam J. Makarucha, Roberto de la Rica, Irene Yarovsky, Molly M. StevensHIV-1 protease is a key enzyme in
the life cycle of HIV/AIDS, as
it is responsible for the formation of the mature virus particle.
We demonstrate here that phage-display peptides raised against this
enzyme can be used as peptide sensors for the detection of HIV-1 protease
in a simple, one-pot assay. The presence of the enzyme is detected
through an energy transfer between two peptide sensors when simultaneously
complexed with the target protein. The multivalent nature of this
assay increases the specificity of the detection by requiring all
molecules to be interacting in order for there to be a FRET signal.
We also perform molecular dynamics simulations to explore the interaction
between the protease and the peptides in order to guide the design
of these peptide sensors and to understand the mechanisms which cause
these simultaneous binding events. This approach aims to facilitate
the development of new assays for enzymes that are not dependent on
the cleavage of a substrate and do not require multiple washing steps.