This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This is a program proposal for beamtime at APS to further several projects underway in our laboratory, including studies of nitric oxide transport proteins called nitrophorins, a multi-copper oxidase, CueO, and a DNA binding protein, NM23-H2. Our work is currently supported by two NIH grants. Four additional proposals are pending. We have determined the structures of nitrophorins from two blood-sucking insects, Rhodnius prolixus (rNP) and Cimex lectularius (cNP). All nitrophorins contain a highly-ruffled ferric heme. Using APS beamline 14-BM-C, we have obtained 0.75 A data on rNP4, which is allowing us to determine heme distortion and ligand conformation in exquisite detail. Diffraction measurements of nitrophorin-NO complexes must be conducted at a synchrotron source since photoreduction occurs when CuKa radiation is used. Laue experiments to delineate changes occurring upon NO dissociation are also planned. CueO is a multicopper oxidase from E. coli expressed under conditions of copper stress. Crystals of CueO diffract to 1.2 A, - considerably better than other multi-copper oxidases. Consequently, we believe CueO is a good candidate for experiments aimed at determining the structure of catalytic intermediates, either by trapping or time-resolved studies. NM23-H2 is a nucleotide-diphosphate kinase that has been shown to promote c-myc (oncogenic) expression. Recently it has been proposed that NM23-H2 binding to DNA from the c-myc promoter region stimulates c-myc expression. We have crystallized native and mutant NM23-H2 in four forms and have begun to study oligonucleotide binding to NM23-H2. Both NM23-H2 and a catalytically deficient mutant diffract to better than 2.5 A in-house, and we have solved structures in two crystal forms by molecular replacement (Rcryst = 0.21). As part of this project, we plan to investigate the structures of putative G-quadruplexes and I-motifs formed by oligonucleotides with sequences from the promoter region of c-myc.
| Weingarten, Adam S; Dannenhoffer, Adam J; Kazantsev, Roman V et al. (2018) Chromophore Dipole Directs Morphology and Photocatalytic Hydrogen Generation. J Am Chem Soc 140:4965-4968 |
| Yang, Cheolhee; Choi, Minseo; Kim, Jong Goo et al. (2018) Protein Structural Dynamics of Wild-Type and Mutant Homodimeric Hemoglobin Studied by Time-Resolved X-Ray Solution Scattering. Int J Mol Sci 19: |
| Kazantsev, Roman V; Dannenhoffer, Adam J; Weingarten, Adam S et al. (2017) Crystal-Phase Transitions and Photocatalysis in Supramolecular Scaffolds. J Am Chem Soc 139:6120-6127 |
| Fournier, Bertrand; Sokolow, Jesse; Coppens, Philip (2016) Analysis of multicrystal pump-probe data sets. II. Scaling of ratio data sets. Acta Crystallogr A Found Adv 72:250-60 |
| Cho, Hyun Sun; Schotte, Friedrich; Dashdorj, Naranbaatar et al. (2016) Picosecond Photobiology: Watching a Signaling Protein Function in Real Time via Time-Resolved Small- and Wide-Angle X-ray Scattering. J Am Chem Soc 138:8815-23 |
| Pande, Kanupriya; Hutchison, Christopher D M; Groenhof, Gerrit et al. (2016) Femtosecond structural dynamics drives the trans/cis isomerization in photoactive yellow protein. Science 352:725-9 |
| Weingarten, Adam S; Kazantsev, Roman V; Palmer, Liam C et al. (2015) Supramolecular Packing Controls H? Photocatalysis in Chromophore Amphiphile Hydrogels. J Am Chem Soc 137:15241-6 |
| Pfoh, Roland; Pai, Emil F; Saridakis, Vivian (2015) Nicotinamide mononucleotide adenylyltransferase displays alternate binding modes for nicotinamide nucleotides. Acta Crystallogr D Biol Crystallogr 71:2032-9 |
| Mariette, Céline; Guérin, Laurent; Rabiller, Philippe et al. (2015) The creation of modulated monoclinic aperiodic composites in n-alkane/urea compounds. Z Kristallogr Cryst Mater 230:5-11 |
| Yang, Xiaojing; Stojkovi?, Emina A; Ozarowski, Wesley B et al. (2015) Light Signaling Mechanism of Two Tandem Bacteriophytochromes. Structure 23:1179-89 |
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