X-ray crystallography and SAXS

Complementing the expertise of our X-ray Core, described below, is Dr. Peter Cherepanov, at the London Research Institute. Dr. Cherepanov, with Dr. Alan Engelman, another PCHPI collaborator, solved the initial co-crystal structure of HIV-1 integrase with LEDGF, and subsequently solved the prototype foamy virus intasome. As a member of the PCHPI, Drs Cherepanov and Engelman will work to understand CA protein interactions with cellular components.

Crystal Screening, Crystal Structure and SAXS Core

Director: Angela M. Gronenborn, with Ying Wu as manager.

The Crystal Screening and Crystal Structure core is responsible for 1) refining crystallization and additive screens for protein complexes to optimize the probability of obtaining diffracting crystals, 2) solving structures of HIV and/or host proteins using high-throughput approaches for crystallization, data collection, and structure determination, and 3) carrying out SAXS analysis of proteins, with collaborative input from Dr. Yun-Xing Wang.

The PCHPI makes use of the X-ray facility of the Department of Structural Biology, which occupies ~1500 sq. ft. suite, on the first floor of BST3. The facility is partitioned into six rooms, containing multiple generators and detectors. The range of equipment available allows for optimal data collection from very small and normal sized crystals, and from crystals with large unit cells which typically contain very large proteins, viruses, or protein-protein assemblies. One of the generators also utilizes an ACTOR robot for automatically mounting and collecting data on a series of crystals, rapidly screening them for diffraction characteristics. Three additional labs, devoted to crystallization and computational analysis, contain equipment for automatic preparation of crystallization screening trays and for automatic and unattended imaging of these trays, as well as equipment for microscopy, and data analysis. The X-ray suite is supplemented by two environmentally controlled rooms dedicated to grow, store, and monitor crystals, at ambient temperature and at 4°C.

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