CHAPTER 7
Protein Structure and Dynamics by NMR in Solution
B. BERSCH i , M. BLACKLEDGE i , B. BRUTSCHER i , F. CORDIER!, J.-CHR. Hus i
and D. MARION i
1
Introduction
Due to the genome project, a tremendous number of protein sequences have
already been obtained, which will continue to increase over the next few years. It
is clear from a biological point of view, that this sequential information has to be
related to protein function. This can partially be obtained by careful analysis of
the sequential data, but unfortunately, functional homology is not always
reflected as sequential homology. Another very important connection between
sequence and function is the molecular structure of a protein. Structural information on the atomic level can currently be obtained from X-ray crystallography,
nuclear magnetic resonance (NMR) as well as, under certain circumstances
(sequential homology to a protein with known 3D structure), molecular modelling. The contribution of structural biology to the classification and understanding of protein structures is reflected in the ever growing number of molecular
coordinates deposited in the Brookhaven Protein Databank (pdb).
Since the introduction of the two-dimensional NMR techniques in the 1970s,
many protein structures have been solved using solution NMR spectroscopy. This
technique has constantly evolved with the increase of magnetic field strength and
computer performance, with the introduction of new, i3N_ and l3C edited multidimensional experiments and, very recently, by taking advantage of phenomena
well known in solid state NMR.
One very important feature of high resolution NMR is that the molecule of
interest is studied in solution, thus allowing the characterization of its dynamic
properties under conditions close to those found in vivo. In addition, NMR is a
sensitive probe for local conformational and/or dynamical variations occuring in
a molecule due to ligand-binding, protein-protein interactions or site directed
mutagenesis.
In this chapter we will give an informal introduction to modern biomolecular
NMR, starting with the necessary information required to initiate an NMR study
of a biomolecule: the suitability of a molecule for an NMR investigation and a
description of the experimental conditions. The experimental parameters and
their possible interpretations will be briefly described before we give some more
I Laboratoire de RMN, Institut de Biologie Structurale - Jean Pierre Ebel, CEA-CNRS, 41 rue Jules
Horowitz, 38027 Grenoble, France.
Protein Structure and Dynamics by NMR in Solution
B. BERSCH i , M. BLACKLEDGE i , B. BRUTSCHER i , F. CORDIER!, J.-CHR. Hus i
and D. MARION i
1
Introduction
Due to the genome project, a tremendous number of protein sequences have
already been obtained, which will continue to increase over the next few years. It
is clear from a biological point of view, that this sequential information has to be
related to protein function. This can partially be obtained by careful analysis of
the sequential data, but unfortunately, functional homology is not always
reflected as sequential homology. Another very important connection between
sequence and function is the molecular structure of a protein. Structural information on the atomic level can currently be obtained from X-ray crystallography,
nuclear magnetic resonance (NMR) as well as, under certain circumstances
(sequential homology to a protein with known 3D structure), molecular modelling. The contribution of structural biology to the classification and understanding of protein structures is reflected in the ever growing number of molecular
coordinates deposited in the Brookhaven Protein Databank (pdb).
Since the introduction of the two-dimensional NMR techniques in the 1970s,
many protein structures have been solved using solution NMR spectroscopy. This
technique has constantly evolved with the increase of magnetic field strength and
computer performance, with the introduction of new, i3N_ and l3C edited multidimensional experiments and, very recently, by taking advantage of phenomena
well known in solid state NMR.
One very important feature of high resolution NMR is that the molecule of
interest is studied in solution, thus allowing the characterization of its dynamic
properties under conditions close to those found in vivo. In addition, NMR is a
sensitive probe for local conformational and/or dynamical variations occuring in
a molecule due to ligand-binding, protein-protein interactions or site directed
mutagenesis.
In this chapter we will give an informal introduction to modern biomolecular
NMR, starting with the necessary information required to initiate an NMR study
of a biomolecule: the suitability of a molecule for an NMR investigation and a
description of the experimental conditions. The experimental parameters and
their possible interpretations will be briefly described before we give some more
I Laboratoire de RMN, Institut de Biologie Structurale - Jean Pierre Ebel, CEA-CNRS, 41 rue Jules
Horowitz, 38027 Grenoble, France.