Solution and Solid-state NMR spectroscopy
The PCHPI counts three world-renowned NMR spectroscopists among its membership. Dr. Angela M. Gronenborn specializes in solution NMR spectroscopy, having contributed to the development of the method, with particular expertise in the analysis of relatively large proteins and protein complexes. Dr. Rieko Ishima is a leader in the development of NMR relaxation methods to understand protein dynamics in solution, with particular emphasis on the RNase domain of HIV-1 reverse transcriptase. Dr. Tatyana Polenova, who applies solid-state NMR techniques to understand protein assemblies, is focused on developing and applying ultra-high field solid-state NMR methods to understand the capsid of HIV-1. The NMR core of the PCHPI is anchored by the technical expertise of Dr. In-Ja Byeon, who has made significant contributions to the field, in several areas with work on the capsid protein (CA), APOBEC proteins, and HIV-1 Nef.
NMR Screening and Solution Structures Core (NMR Core)
Director: Angela M. Gronenborn, with In-Ja L. Byeon as manager.
The NMR Screening and Solution Structure core is responsible for (1) assessing the structural properties of HIV proteins to determine their suitability for further structural study, (2) screening potential binding partners, provided by collaborators, for putative interactions with HIV proteins, and (3) structure determination of interacting complexes that have been identified through screening.
The NMR Screening and Solution Structure Core completes structural work on purified biological macromolecules and complexes (up to 80kDa) in aqueous solution. This material, produced by the Protein Production Core is, typically, uniformly 15N labeled or 15N/13C/D labeled, proteins and/or nucleic acids. Sample volumes are typically 250-500 ul and experiments are performed at temperatures and solution conditions suitable for ensuring structural and functional integrity of the molecules. In addition to solution NMR spectroscopy, two instruments (wide-bore 600 MHz and 800 MHz) have capabilities for solid state NMR, allowing us to perform work on membrane proteins.
The PCHPI makes use of the NMR facility of the Department of Structural Biology that occupies ~10,000 sq. ft. of specifically designed space in the basement of the BST3, where a complement of high-field spectrometers is housed. Overall, the NMR facility is extremely well equipped for all types of biomolecular NMR and currently has six five-channel Bruker AVANCE spectrometers, three operating at 600MHz (including one wide-bore magnet), one each at 700MHz, 800MHz, and 900MHz. All ultra shielded magnets are equipped with cryoprobes for increased sensitivity.