The recent development of cryoprobe technology is providing tremendous gains in the inherent sensitivity of solution-state NMR spectrometers. This achievement is of enormous value for NMR studies in the field of structural biology, due to the fundamental role of instrument sensitivity in determining the feasibility of most biomolecular NMR research proposals. Current cryoprobe technology has allowed sensitivity improvements by factors in the range of 2 to 4 to be realized, with the exact gain depending on the conductivity of the sample buffer. In contrast, improvements in sensitivity of conventional NMR probes by as little as 20% have been exceptionally difficult to achieve, and would be considered quite significant. Utilization of a cryoprobe allows sample concentrations to be reduced by factors of 2-4, which greatly facilitates work on samples with low solubility, a tendency to aggregate, or in cases where the sample material is especially difficult and/or costly to obtain. Alternatively, it is possible, by using a cryoprobe, to reduce total data acquisition times by factors of 4 to 16, which is extremely valuable when working on samples of limited stability, when considering projects which require extensive amounts of data to be collected, for allowing for the possibility of performing low-sensitivity experiments in realistic periods of time, and for significantly improving the overall throughput on very costly, ultra-high field NMR spectrometers. In order for structural biologists to continue to tackle increasingly difficult problems, it is vital to exploit whatever technological advances are available. The current proposal requests funds to purchase a triple resonance 5 mm1H-{13C,15N} pulsed field gradient cryoprobe and required accessories for the 800 MHz NMR spectrometer that is housed in the structural biology/NMR facility within the University of Cincinnati College of Medicine. This facility supports a large number of NIH-funded investigators, and acquisition of the cryoprobe will greatly facilitate the existing research projects and will allow new and even more challenging problems to be tackled.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biomedical Research Support Shared Instrumentation Grants (S10)
Project #
1S10RR019077-01
Application #
6732309
Study Section
Special Emphasis Panel (ZRG1-PB (30))
Program Officer
Tingle, Marjorie
Project Start
2004-05-01
Project End
2005-12-31
Budget Start
2004-05-01
Budget End
2005-12-31
Support Year
1
Fiscal Year
2004
Total Cost
$345,443
Indirect Cost
Name
University of Cincinnati
Department
Genetics
Type
Schools of Medicine
DUNS #
041064767
City
Cincinnati
State
OH
Country
United States
Zip Code
45221
Kojetin, Douglas J; Matta-Camacho, Edna; Hughes, Travis S et al. (2015) Structural mechanism for signal transduction in RXR nuclear receptor heterodimers. Nat Commun 6:8013
Liu, Sheng; Decker, Aaron; Howell, Mike et al. (2013) ¹H, ¹³C and ¹?N resonance assignment of the N-terminal domain of human lysyl aminoacyl tRNA synthetase. Biomol NMR Assign 7:289-92
Kuo, Shiu-Ming; Wang, Li-Yuan; Yu, Siyuan et al. (2013) The N-terminal basolateral targeting signal unlikely acts alone in the differential trafficking of membrane transporters in MDCK cells. Biochemistry 52:5103-5116
Liu, Sheng; Howell, Michael; Melby, Joel et al. (2012) 1H, 13C and 15N resonance assignment of the anticodon binding domain of human lysyl aminoacyl tRNA synthetase. Biomol NMR Assign 6:173-6
Doerdelmann, Thomas; Kojetin, Douglas J; Baird-Titus, Jamie M et al. (2012) Structural and biophysical insights into the ligand-free Pitx2 homeodomain and a ring dermoid of the cornea inducing homeodomain mutant. Biochemistry 51:665-76
Hughes, Travis S; Chalmers, Michael J; Novick, Scott et al. (2012) Ligand and receptor dynamics contribute to the mechanism of graded PPAR? agonism. Structure 20:139-50
Doerdelmann, Thomas; Kojetin, Douglas J; Baird-Titus, Jamie M et al. (2012) ¹H, ¹³C and ¹?N chemical shift assignments for the human Pitx2 homeodomain in complex with a 22-base hairpin DNA. Biomol NMR Assign 6:79-81
Doerdelmann, Thomas; Kojetin, Douglas J; Baird-Titus, Jamie M et al. (2011) 1H, 13C and 15N chemical shift assignments for the human Pitx2 homeodomain and a R24H homeodomain mutant. Biomol NMR Assign 5:105-7
Refaei, Mary Anne; Combs, Al; Kojetin, Douglas J et al. (2011) Observing selected domains in multi-domain proteins via sortase-mediated ligation and NMR spectroscopy. J Biomol NMR 49:3-7