title: Effects of PRE and POST therapy drug-pressure selected mutations on HIV-1 protease conformational sampling.
abstract: Conformational sampling of pre- and post-therapy subtype B HIV-1 protease sequences derived from a pediatric subject infected via maternal transmission with HIV-1 were characterized by double electron-electron resonance spectroscopy. The conformational ensemble of the PRE construct resembles native-like inhibitor bound states. In contrast, the POST construct, which contains accumulated drug-pressure selected mutations, has a predominantly semi-open conformational ensemble, with increased populations of open-like states. The single point mutant L63P, which is contained in PRE and POST, has decreased dynamics, particularly in the flap region, and also displays a closed-like conformation of inhibitor-bound states. These findings support our hypothesis that secondary mutations accumulate in HIV-1 protease to shift conformational sampling to stabilize open-like conformations, while maintaining the predominant semi-open conformation for activity.
authors.affiliation: Department of Chemistry, University of Florida, Gainesville, FL 32611-7200, USA.
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Carter
authors.affiliation: Department of Chemistry, University of Florida, Gainesville, FL 32611-7200, USA.
authors.forename: Estrella G
authors.initials: EG
authors.lastname: Gonzales
authors.affiliation: Department of Chemistry, University of Florida, Gainesville, FL 32611-7200, USA.
authors.forename: Xi
authors.initials: X
authors.lastname: Huang
authors.affiliation: Department of Chemistry, University of Florida, Gainesville, FL 32611-7200, USA.
authors.forename: Adam N
authors.initials: AN
authors.lastname: Smith
authors.affiliation: Department of Chemistry, University of Florida, Gainesville, FL 32611-7200, USA.
authors.forename: Ian Mitchelle S
authors.initials: IM
authors.lastname: de Vera
authors.affiliation: Department of Chemistry, University of Florida, Gainesville, FL 32611-7200, USA.
authors.forename: Peter W
authors.initials: PW
authors.lastname: D'Amore
authors.affiliation: Advanced Magnetic Resonance Imaging and Spectroscopy Facility, McKnight Brain Institute, University of Florida, Gainesville, FL 32610, USA.
authors.forename: James R
authors.initials: JR
authors.lastname: Rocca
authors.affiliation: Department of Pathology, Immunology and Laboratory Medicine, University of Florida College of Medicine, Gainesville, FL 32610-3633, USA.
authors.forename: Maureen M
authors.initials: MM
authors.lastname: Goodenow
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Florida College of Medicine, Gainesville, FL 32610-0245, USA.
authors.forename: Ben M
authors.initials: BM
authors.lastname: Dunn
authors.affiliation: Department of Chemistry, University of Florida, Gainesville, FL 32611-7200, USA. Electronic address: fanucci@chem.ufl.edu.
authors.forename: Gail E
authors.initials: GE
authors.lastname: Fanucci
mesh_terms: D019380:Anti-HIV Agents / Q000494:pharmacology*; D024882:Drug Resistance, Viral / Q000235:genetics; D016333:HIV Protease / Q000737:chemistry* / Q000235:genetics*; D015497:HIV-1 / Q000187:drug effects* / Q000201:enzymology* / Q000235:genetics; D008958:Models, Molecular; D009154:Mutation; D017354:Point Mutation; D011487:Protein Conformation; D013997:Time Factors
chemical_list: D019380:Anti-HIV Agents; D016333:HIV Protease
keywords: Double electron–electron resonance; HIV-1; Protease; Pulsed electron double resonance; Site-directed spin labeling; Spin-labeling