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. Uridine 5?-diphospho -N-acetylglucosamine (UDP-GlcNAc) is an essential donor for synthesis of a diverse array of glycans &glycoconjugates used in fields spanning from therapeutic medicine and vaccines to food industry. In nature, various organisms employ different and mostly complicated pathways to synthesize UDP-GlcNAc for biosynthesis of cell walls, glycosaminoglycans such as hyaluronic acid, etc. The chemical synthesis of UDP-GlcNAc is also complicated and costly. The final goal of our project is to engineer a simple, economic and efficient enzymatic synthesis of UDP-GlcNAc. In order to accomplish this we are using a two enzyme system with GlcNAc and ATP as starting material. GlcNAc is converted to GlcNAc-1-P by an enzyme called NahK while utilizing ATP. GlcNAc-1-P is further converted to UDP-GlcNAc by GlmU while utilizing UTP. Whereas GlmU has been characterized structurally and biochemically, NahK is not. The specific focus of current project is to investigate the structure of NahK and study its binding with various substrates and substrate analogs. It is hoped that these studies will eventually lead to development of a more stable and efficient enzyme with optimum substrate specificity. NahK (B.longum) is a soluble acidic enzyme of moderate molecular weight (~ 40 kD). The protein belongs to protein kinase superfamily &functions as N-acetylhexosamine kinase. It is a relatively novel and unique enzyme in being the first reported enzyme that acts as a gluco-type kinase (ref. #1) i.e., can phosphorylate glucose and its analogs. Crystallization attempts were undertaken to obtain structure of this enzyme. Small crystals of 40-50 microns in largest dimension could finally be obtained. Being too small they could diffract only poorly at the home X-ray source (5 angstrom resolution). Selenomethionine containing protein was obtained by using methionine auxotrophic host strain. The crystals obtained from this Se-protein would be tried for MAD studies.
| 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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