Quantitative assessment of in vivo HIV protease activity using genetically engineered QD-based FRET probes.
HIV protease plays a central role in its life cycle leading to release of functional viral particles. It has been successfully used as a therapeutic target to block HIV infection. Several protease inhibitors (PIs) are currently being employed as a part of anti-HIV therapy. However, the constant genetic drift in the virus leads to accumulation of mutations in both cleavage site and the protease, resulting in resistance and failure of therapy. We reported the use of a quantum dot (QD)-based protein probe for the in vivo monitoring of HIV-1 protease activity based on fluorescence resonance energy transfer. In the current study, we demonstrate the utility of this approach by quantifying the in vivo cleavage rates of three known protease and cleavage site mutations in the presence or absence of different PIs. The changes in IC50 values for the different PIs were similar to that observed in patients, validating our assay as a rapid platform for PI screening.
Department of Chemical and Environmental Engineering, University of California, Riverside, California.
Lakshmi N
LN
Cella
Payal
P
Biswas
Marylynn V
MV
Yates
Ashok
A
Mulchandani
Wilfred
W
Chen
D031541:Fluorescence Resonance Energy Transfer; D016333:HIV Protease / Q000378:metabolism*; D017320:HIV Protease Inhibitors / Q000494:pharmacology*; D015497:HIV-1 / Q000201:enzymology*; D045663:Quantum Dots
D017320:HIV Protease Inhibitors; D016333:HIV Protease
FRET; HIV protease; in vivo kinetics; quantum dots; viral detection