Premium
Quantitative assessment of in vivo HIV protease activity using genetically engineered QD‐based FRET probes
Author(s) -
Cella Lakshmi N.,
Biswas Payal,
Yates Marylynn V.,
Mulchandani Ashok,
Chen Wilfred
Publication year - 2014
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.25199
Subject(s) - protease , förster resonance energy transfer , in vivo , cleavage (geology) , hiv 1 protease , human immunodeficiency virus (hiv) , chemistry , virus , green fluorescent protein , in vitro , biophysics , microbiology and biotechnology , biology , virology , fluorescence , biochemistry , enzyme , genetics , gene , paleontology , physics , quantum mechanics , fracture (geology)
ABSTRACT 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 IC 50 values for the different PIs were similar to that observed in patients, validating our assay as a rapid platform for PI screening. Biotechnol. Bioeng. 2014;111: 1082–1087. © 2014 Wiley Periodicals, Inc.