NMR spectroscopy is a vital tool for biologists that aids in the structural determination, dynamics, and interactions of biological macromolecules, especially proteins. It offers advantages over traditional methods like x-ray crystallography due to its sensitivity to minute changes and noninvasiveness, allowing analysis in physiological environments and living cells. A variety of NMR methods exist to derive structural restraints essential for protein analysis, including the nuclear overhauser effect (NOE) for assessing structural integrity in complex environments.

1. NMR spectroscopy
BY
JYOTI REKHA PATRA

2. • NMR spectroscopy is a powerful tool for biologists interested in the
structure, dynamics, and interactions of biological macromolecules.
• It is a routine method for the structural determination of
proteins and protein complexes at atomic resolution and
can also provide rich information regarding both
conformational and interactional dynamics occurring on
time scales ranging from picoseconds to seconds or even
days and in various sample states ranging from dilute
solutions to living cells.

3. PROCEDURE:-
ISOTOPE LABELLED PROTEIN SAMPLE PREPARATION
DATA ANALYSIS OF NMR, PARTICULARLY ASSIGNING H1, N15,C13 ATOMS IN PROTEIN MOLECULE
STRUCTURAL QUALITY ASSESMENT
STRUCTURAL CALCULATION USING DISTANCE & ORIENTATION

4. • NMR analysis of
proteins is much less
dependent on sample
conditions compared
to X-ray or cryo-EM
methods and is highly
sensitive to subtle
changes in
chemical environment
s
one thing, as a noninvasive method, NMR
is ideal for protein analysis not only in
buffers mimicking physiological
environment but also in actual living cells.
It is also feasible to analyze membrane
protein structures in close-to-native
environments by using both solid-state
and solution NMR.
For another thing, a variety of NMR
methods have been developed to obtain
information regarding protein dynamics.

5. Basic
Concepts in
Protein NMR
Protein structural analysis by NMR relies on the
collection of experimental structural restraints,
which typically include the NOE-derived local
distance restraints (≤6 Å), hydrogen bonds, and
dihedral angle restraints.
• Nuclear Overhauser Effect (NOE)
NOE is one of the most important phenomena in
NMR spectroscopy

6. Structural
Analysis of
Proteins in
Complex
Environments
Most protein structures are
determined either in a simple buffer
by NMR or in a crystal form by X-ray
crystallography.
For protein structural determination
by NMR in a dilute solution, diverse
choices of strategies have been
developed to cope with situations of
different complexities.

9. THANK YOU