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Biomolecular NMR Core Facility

Investigators
Prakash, Om
Institutions
Kansas State University
Start date
2009
End date
2014
Objective
  1. This Biomolecular nuclear magnetic resonance laboratory will assist KAES researchers in conducting nationally competitive research at Kansas State University in diverse areas of agriculture biochemistry, structure biology and in characterization of chemicals and their products in clinical, food, industrial, environmental and agriculture products samples.
  2. Another objective of this NMR facility at K-State will be to initiate new NMR-linked grant proposals with KAES researchers, developing cutting-edge techniques and make sure that our students are educated and trained in using latest multinuclear, multi-dimensional NMR techniques.
  3. The Biomolecular NMR laboratory will provide and continue services of an expensive NMR instrumentation and solution structure determination infrastructure to KAES investigators which is difficult to acquire, operate and maintain by an individual investigator.
  4. This laboratory will also support KAES investigators in molecular modeling and dynamics analyses of NMR experimental data. These analyses will guide the design of future experiments, particularly those employing recombinant DNA technology to create new protein structures useful in developing resistance to insects and diseases in plants and animals, designing crops and animals with specific properties for special end uses and other useful agricultural commodities.
More information
Non-Technical Summary: a) High-resolution solution NMR studies of proteins/protein domains and other macromolecules will provide valuable information to resolve many structural components and to quantify the changes produced in proteins or other molecules by chemical process. This technology will enhance our understanding at the molecular level about the three dimensional structure and folding/unfolding behavior of various proteins which in turn will explain processes such as formation of the large quaternary protein structure and their function. b) The biomolecular NMR facility will obtain scientific information, generate preliminary data and provide support letters that will be included in the grant applications of KAES investigators for extramural funding. The availability of 500 MHz NMR spectrometer and more than ten work stations equipped with faster microprocessors will provide hands-on experience to researchers and students with multi-dimensional NMR data acquisition and analysis. The lab is capable of performing modern multidimensional NMR analyses to determine the following: three-dimensional structure of carbohydrates, peptides, soluble proteins; protein dynamics; structural and reactive nature of metal-protein complexes; protein folding; protein interactions with other proteins, lipids, or ligands; nucleic acid structure and dynamics; identification of metabolites and other biological molecules. The major impact of this state-of-the-art laboratory to the KAES investigators and educators will be the advanced level of educational training and research performed at K-State. This will also help in recruiting potential graduate students and new faculty. c) The expected outcome of NMR data based molecular modeling and dynamic analysis of proteins, peptides and other biological macromolecules will be to advance our knowledge on molecular recognition, complex formation, conformational equilibria, folding and intra- as well as intermolecular interactions. This information will assist KAES researcher in designing new proteins and agricultural products with specific properties for special end uses. The engineered proteins can be patented and will have the potential to promote a long term economic growth in Kansas. It is also noteworthy that this facility is one of the few NMR centers in the Midwest conducting structure and dynamics studies of peptides, proteins and other biomolecules. Prior to establishing this laboratory, KAES researchers were dependent on out-of-state facilities.

Approach: Using a recently upgraded 500 MHz Varian digital NMR spectrometer and using either a 3mM triple resonance NMR nano-probe or micro-capillary triple resonance probe as required, the Biomolecular NMR Laboratory will perform NMR experiments for macromolecule dynamics studies, protein NMR studies, and NMR structural studies on phosphorylated wheat starch. These studies will require the use of well established protocols using 1-D, 2-D, and 3-D experimental techniques to include: 2-D homonuclear spin correlated spectroscopy (COSY), 2-D total correlation spectroscopy (TOCSY), 2D proton - proton nuclear Overhauser effect spectroscopy (NOESY), rotating frame Overhauser effect spectroscopy (ROESY), carbon-13 and nitrogen-15 NMR relaxation measurements and hydrogen exchange kinetics experiments, difference spectroscopy and heteronuclear single quantum coherence (HSQC), and triple resonance experiments including CBCA(CO)NH, CA(CO)NH, H(CA)(CO)NH and HNCACB. All data recieved will be analyzed and graphically modeled using the software modules: QUANTA/X-PLOR, CNS, MOLMOL and PyMOL which are designed for structure calculation and molecular graphics modeling. The results of NMR studies carried out by KAES investigators will be used for grant proposals, meeting presentations and for peer reviewed research publications.

Funding Source
Nat'l. Inst. of Food and Agriculture
Project source
View this project
Project number
KS421
Accession number
217493
Categories
Microbiome