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. Our crystallographic studies of benzoylformate decarboxylase (BFD), up to 1A resolution, suggest the mode of substrate binding and the mechanism of catalysis. A continuation of this study promises future lessons. The results emphasize that each reside in the active site plays more than one role in catalysis. To identify atoms that are unexpectedly bound to the ThDP under certain conditions, we plan to collect MAD or SAD data at the absorption edges of various elements that may be present. Time-resolved crystallography, both through Laue diffraction and mutant structures, will allow the observation of reaction steps as well as theroles of the residues in these steps. We also hope to solve structures of other family members. The fold of BFD is similar to that of other enzymes that bind thiamine diphosphate, and the residues that bind the cofactor are conserved. Surprisingly, other potential catalyticresidues are not conserved. These results underscore the prime importance of the cofactor in carrying out the underlying chemistry required for the enzyme-catalyzed reaction. BFD providesat least nine crystal forms that bind different compounds, allowing us to visualize the courseof the reaction. Further, the phenyl ring of the substrate, benzoylformate, allows the observation of intermediates through spectroscopy. The results in our experiments have suggested that the decarboxylation results in a bicarbonate in the active site, with a serine in the active site involved in its movement. The power of the active site was demonstrated bythe ability of benzoylphosphonate to phosphorylate the serine. A strong carbon-phosphate bond such as in benzoylphosphonate has never been seen to phosphorylate a serine. In addition, mutation of the serine, glutamate or histidine in the active site to alanine can cause oxidation of the thiamin diphosphate. This suggests that the enzyme is composed not only to form the product, but also to prevent errors on the way.
| 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 |
Showing the most recent 10 out of 120 publications
