The primary rationale for the PCHPI organization is to maximize interdisciplinary collaboration and access to the PCHPI resources, while maintaining focused expertise in technical disciplines. 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