Chemistry Reference
In-Depth Information
NMR Studies of Metalloproteins
Hongyan Li and Hongzhe Sun
Abstract Metalloproteins represent a large share of the proteomes, with the intrinsic
metal ions providing catalytic, regulatory, and structural roles critical to protein
functions. Structural characterization of metalloproteins and identification of metal
coordination features including numbers and types of ligands and metal-ligand geom-
etry, and mapping the structural and dynamic changes upon metal binding
are significant for understanding biological functions of metalloproteins. NMR spec-
troscopy has long been used as an invaluable tool for structure and dynamic studies of
macromolecules. Here we focus on the application of NMR spectroscopy in character-
ization of metalloproteins, including structural studies and identification of metal
coordination spheres by hetero-/homo-nuclear metal NMR spectroscopy. Paramag-
netic NMR as well as
13
C directly detected protonless NMR spectroscopy will also be
addressed for application to paramagnetic metalloproteins. Moreover, these techniques
offer great potential for studies of other non-metal binding macromolecules.
Keywords C direct detection
Metal coordination
Metalloprotein
NMR
spectroscopy
Paramagnetic metalloprotein
Contents
1
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
2
The Contribution of NMR to Structural Metalloproteins ................................... 71
2.1 Conventional Method for Structure Determination of Metalloproteins . . ............ 71
2.2 Utilization of Chemical Shifts to Generate Structures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76
3
Identification of Metal Coordination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76
3.1 Homonuclear and Heteronuclear Metal NMR Spectroscopy ......................... 76
3.2 Chemical Shift Perturbation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82
