# X-ray Screening and Rapid Structure Determination

> **NIH NIH P50** · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · 2020 · $199,786

## Abstract

The aim of the Crystallography Core is to provide robotic crystallization and screening for all projects that aim
to crystallize HIV-host protein complexes for structure determination involving HIV-1 Vif, Vpu, Nef, Tat and Rev
(Projects 1, 3, 4, 5 and 6). A second function is to provide assistance during on-site, or remote access X-ray
data collection, or entirely ‘Fed-Ex’ access to X-ray diffraction at Beamline 8.3.1, and solution scattering access
at Beamline 12.3.1 at the Advanced Light Source (ALS) in Berkeley. Data frames can be reduced and
structures determined on-site. A third function is to provide for structure refinement as necessary so that a
biochemist can efficiently go through all steps without any knowledge of the crystallographic process.
Investigators who want to learn or direct the crystallography will have access to any training necessary, without
restriction. The process of crystallization is facilitated by two, two-nanoliter drop setting robots (TTP Mosquitos
– one with capacity to set up LCP experiments) that set up crystallization trials, generally in 96 well plate based
systems, at both 4°C and at 20°C. The trays are incubated at either 4°C or 20°C in a second type of robotic
system. These systems are capable of storing a thousand trays and visualizing them on a programmed
schedule (Formulatrix Rock Imager) with advanced optics, including crossed-polarization and UV screening
capacity to validate protein crystals. The images are stored for remote screening online. Once crystals are
available, the Core provides for screening of crystals at the Advanced Light Source (ALS) in Berkeley at
Beamlines 8.3.1. or 12.3.1. The system allows screening many crystal loops robotically, and returning to collect
datasets on selected screening hits. Beamline 12.3.1 allows for completely remote data collection, and also for
running in a small- or wide-angle X-ray scattering mode (SAXS, WAXS) for solution scattering. The systems at
ALS are maintained and evolved to be forefront technology for the vast majority of protein complex crystals.
This includes ultra flat focusing optics and a new Pilatus3 6M detector. Data processing can be accomplished
in parallel with data collection. An experienced two member staff is available at the beamline so that any HARC
Center members may attend to use the system, returning with reduced data and/or a structure determined by
anomalous diffraction or molecular replacement, as appropriate. Anomalous diffraction using selenium
substitution or endogenous sulfur is feasible. The system has given rise to the vast majority of HIV related
structures from members of the HARC Center Project teams. An experienced investigator is available for
guidance or execution of refinement. The system can be used by biochemists who have no experience with
crystallography, or can be used hands-on at each step as matched to investigator strengths. On-site training is
provided at every step as needed. The systems are developed and ma...

## Key facts

- **NIH application ID:** 9993239
- **Project number:** 5P50AI150476-14
- **Recipient organization:** UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
- **Principal Investigator:** Robert M Stroud
- **Activity code:** P50 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $199,786
- **Award type:** 5
- **Project period:** 2007-08-27 → 2022-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9993239

## Citation

> US National Institutes of Health, RePORTER application 9993239, X-ray Screening and Rapid Structure Determination (5P50AI150476-14). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/9993239. Licensed CC0.

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