During the past year we measured multiwavelength anomalous diffraction (MAD) from cocrystals of the DNA binding domain of Interferon regulatory factor-1 (IRF-1) complexed to DNA. IRF-1 plays a key role in the regulation of interferons and interferon inducible genes. The MAD data were measured at three wavelengths corresponding to the edge (0.92048?) and peak (0.9198?) of the Br K absorption profile, plus a remote point (0.90496?). Because of the R3 symmetry of the cocrystal, the Freidel pairs could only be recorded by the inverse beam method (f and f + 180o). The data were collected by the oscillation method (Df=1o), with no overlap between successive frames. Inverse data were recorded after every 4-6o of rotation. By the end of the data collection the cocrystal had been rotated 260o. The MAD data were successfully processed and used to determine the structure of this biologically important complex. A manuscript describing the structure is currently in progress. We also measured a high resolution data from crystals of the enzyme FokI in its """"""""free"""""""" state. Crystals of free enzyme diffracted to ~ 3.5? at home, but at CHESS we were able to measure data extending to 2.3? resolution. FokI is an unusual restriction endonuclease that has its DNA recognition and cleavage functions located on distinct subdomains. The structure of the free enzyme will complement the structure of the FokI-DNA complex that was determined using data measured at CHESS during the previous year.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR001646-20
Application #
6667770
Study Section
Project Start
2002-09-30
Project End
2003-08-14
Budget Start
Budget End
Support Year
20
Fiscal Year
2002
Total Cost
$142,703
Indirect Cost
Name
Cornell University
Department
Type
DUNS #
City
Ithaca
State
NY
Country
United States
Zip Code
14850
Kozlov, Guennadi; Wong, Kathy; Gehring, Kalle (2018) Crystal structure of the Legionella effector Lem22. Proteins 86:263-267
Ménade, Marie; Kozlov, Guennadi; Trempe, Jean-François et al. (2018) Structures of ubiquitin-like (Ubl) and Hsp90-like domains of sacsin provide insight into pathological mutations. J Biol Chem 293:12832-12842
Xu, Jie; Kozlov, Guennadi; McPherson, Peter S et al. (2018) A PH-like domain of the Rab12 guanine nucleotide exchange factor DENND3 binds actin and is required for autophagy. J Biol Chem 293:4566-4574
Dean, Dexter N; Rana, Pratip; Campbell, Ryan P et al. (2018) Propagation of an A? Dodecamer Strain Involves a Three-Step Mechanism and a Key Intermediate. Biophys J 114:539-549
Chen, Yu Seby; Kozlov, Guennadi; Fakih, Rayan et al. (2018) The cyclic nucleotide-binding homology domain of the integral membrane protein CNNM mediates dimerization and is required for Mg2+ efflux activity. J Biol Chem 293:19998-20007
Xu, Caishuang; Kozlov, Guennadi; Wong, Kathy et al. (2016) Crystal Structure of the Salmonella Typhimurium Effector GtgE. PLoS One 11:e0166643
Cogliati, Massimo; Zani, Alberto; Rickerts, Volker et al. (2016) Multilocus sequence typing analysis reveals that Cryptococcus neoformans var. neoformans is a recombinant population. Fungal Genet Biol 87:22-9
Oot, Rebecca A; Kane, Patricia M; Berry, Edward A et al. (2016) Crystal structure of yeast V1-ATPase in the autoinhibited state. EMBO J 35:1694-706
Lucido, Michael J; Orlando, Benjamin J; Vecchio, Alex J et al. (2016) Crystal Structure of Aspirin-Acetylated Human Cyclooxygenase-2: Insight into the Formation of Products with Reversed Stereochemistry. Biochemistry 55:1226-38
Bauman, Joseph D; Harrison, Jerry Joe E K; Arnold, Eddy (2016) Rapid experimental SAD phasing and hot-spot identification with halogenated fragments. IUCrJ 3:51-60

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