Related Experiment Video
Updated: Jun 21, 2026
11:24
High Precision FRET at Single-molecule Level for Biomolecule Structure Determination
Published on: May 13, 2017
NMR assignments of the binary hvDHFR1:folate complex
Arezue F B Boroujerdi1, Bulent Binbuga, John K Young
1Department of Chemistry, Mississippi State University, Box 9573, Mississippi State, MS 39762, USA.
Biomolecular NMR Assignments
|July 29, 2009
Summary
Nuclear magnetic resonance (NMR) studies of the human dihydrofolate reductase (hDHFR1) enzyme and its folate substrate provide insights into salt
Area of Science:
- Biochemistry and enzymology
- Structural biology
- Biophysical chemistry
Background:
- Enzyme activity is crucial for biological processes.
- Understanding factors influencing enzyme function, such as salt concentration, is essential.
- Human dihydrofolate reductase (hDHFR1) is a key enzyme in folate metabolism.
Purpose of the Study:
- To investigate the impact of salt concentration on enzyme activity.
- To characterize the structural and dynamic properties of the hDHFR1:folate complex.
- To establish a foundation for future studies on salt-dependent enzyme flexibility.
Main Methods:
- Nuclear Magnetic Resonance (NMR) spectroscopy was employed.
- Chemical shift assignments were performed for the 17.9 kDa hDHFR1:folate complex.
- Relaxation studies were initiated to probe enzyme dynamics.
Main Results:
- Successful chemical shift assignments were achieved for the hDHFR1:folate complex.
- These assignments provide a basis for understanding the enzyme's structure.
- The data pave the way for future relaxation studies.
Conclusions:
- NMR studies offer a powerful approach to understanding salt effects on enzyme function.
- The obtained chemical shift assignments are critical for further dynamic investigations.
- This work facilitates future research into salt-induced changes in enzyme flexibility.
More Related Videos
06:21Multi-Gene Single Nucleotide Polymorphism Detection in Gastric Cancer Based on Ion Semiconductor Sequencing Platform
Published on: May 10, 2024
11:27Single-Molecule Förster Resonance Energy Transfer Methods for Real-Time Investigation of the Holliday Junction Resolution by GEN1
Published on: September 18, 2019
Related Concept Videos
01:13¹H NMR: Complex Splitting
A proton M that is coupled to a proton X results in doublet signals for M. However, NMR-active nuclei can be simultaneously coupled to more than one nonequivalent nucleus. When M is coupled to a second proton A, such as in styrene oxide, each peak in the doublet is split into another doublet.
Splitting diagrams or splitting tree diagrams are routinely used to depict such complex couplings. While drawing splitting diagrams, the splitting with the larger coupling constant is usually applied first.
Splitting diagrams or splitting tree diagrams are routinely used to depict such complex couplings. While drawing splitting diagrams, the splitting with the larger coupling constant is usually applied first.
01:192D NMR: Heteronuclear Single-Quantum Correlation Spectroscopy (HSQC)
Heteronuclear single-quantum correlation spectroscopy (HSQC) is a 2D NMR technique that reveals one-bond correlations between hydrogen and a heteronucleus. The HSQC experiment is similar to the heteronuclear correlation experiment (HETCOR) but is more sensitive. In the HSQC spectrum, the proton chemical shift is plotted on the horizontal F2 axis, while the 13C chemical shift is plotted on the vertical F1 axis. The corresponding proton and 13C spectra are also shown. The HSQC contour plot does...
02:42Valence Bond Theory
Coordination compounds and complexes exhibit different colors, geometries, and magnetic behavior, depending on the metal atom/ion and ligands from which they are composed. In an attempt to explain the bonding and structure of coordination complexes, Linus Pauling proposed the valence bond theory, or VBT, using the concepts of hybridization and the overlapping of the atomic orbitals. According to VBT, the central metal atom or ion (Lewis acid) hybridizes to provide empty orbitals of suitable...
01:16Other Nuclides: 31P, 19F, 15N NMR
Many organic, inorganic, and biological molecules contain spin-half nuclei such as nitrogen-15, fluorine-19, and phosphorus-31. As a result, NMR studies of these nuclei have found extensive applications in chemical and biological research.
While fluorine-19 and phosphorous-31 have high natural abundances (100%) and positive gyromagnetic ratios, nitrogen-15 has a low natural abundance and a negative gyromagnetic ratio. However, nitrogen-15 is still preferred over nitrogen-14 (which has a high...
While fluorine-19 and phosphorous-31 have high natural abundances (100%) and positive gyromagnetic ratios, nitrogen-15 has a low natural abundance and a negative gyromagnetic ratio. However, nitrogen-15 is still preferred over nitrogen-14 (which has a high...
01:15¹H NMR of Conformationally Flexible Molecules: Variable-Temperature NMR
The axial and equatorial protons in cyclohexane can be distinguished by performing a variable-temperature NMR experiment. In this process, except for one proton, the remaining eleven protons are replaced by deuterium. The deuterium substitution avoids the possible peak splitting caused by the spin-spin coupling between the adjacent protons. The remaining proton flips between the axial and equatorial positions.
01:09¹H NMR: Pople Notation
The Pople nomenclature system classifies spin systems based on the difference between their chemical shifts. Coupled spins are denoted by capital letters with subscripts indicating the number of equivalent nuclei. When the coupled nuclei have well-separated chemical shifts, they are assigned letters that are far apart in the alphabet, such as A and X. When the difference in chemical shifts is small, coupled nuclei are named using adjacent letters of the alphabet (AB, MN, or XY).
A proton...
A proton...