The Center is organized around four methodological cores (Computational, Cryo-EM/ET, NMR, and HIV Virology) and three scientific projects (1. Maturation; 2. Trafficking, Restriction and Nuclear Entry; and 3. Integration). The cores are complemented by the expertise of our collaborators in several important areas. Close collaboration among the research groups and cores allows the PCHPI to take a “hybrid” approach to answer important structural, biochemical, and biophysical questions about HIV-1.

Computational Core

Derivation of full-scale models that integrate experimental structural data Execution of large-scale and long-timescale simulations Execution of mechanistic studies based on free energy calculations and steered molecular dynamics

 

Cryo-EM & Cryo-ET Core

Cryo-Electron Single Particle Analysis Cryo-Electron Sub-Tomogram Averaging Micro-Electron Diffraction (micro-ED) of nanocrystals and crystal lamellae Cryo-Focused Ion Beam/Scanning EM for cryo-ET and micro-ED of lamellae Cryo-FIB/SEM 3D volume imaging of whole frozen cell/tissue Correlative cryo-flourescence microscopy and cryo-ET (CLEM)

 

NMR Spectroscopy Core

Screening to evaluate the biophysical properties and structural integrity of proteins Determination of structures and dynamics of protein assemblies and their complexes Development of fluorine-based in-cell NMR technology and novel methods for MAS NMR

 

HIV Virology Core

HIV-1 infectivity assays Purification of HIV-1 cores Imaging of early events (Confocal and TIRF microscopy) Immunoblot analysis of HIV-1 particles TEM analysis of HIV-1 particles Quantitative analysis of reverse transcription, nuclear import, and integration HIV-1 integration site analysis