| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
1 Department of Molecular Biology, The Scripps Research Institute, La Jolla, California 92037, USA
2 Department of Physical Chemistry, Lund University, Lund, SE-22100 Sweden
3 Departments of Biochemistry and Physics and Center for Structural Biology, Vanderbilt University, Nashville, Tennessee 37232-0146, USA
Reprint requests to: Dr. Walter J. Chazin, Center for Structural Biology, PRB-896, Vanderbilt University Nashville, TN. 37232-0146; e-mail: walter.chazin{at}vanderbilt.edu; fax: (615) 936-2211.
EF-hand Ca2+-binding proteins participate in both modulation of Ca2+ signals and direct transduction of the ionic signal into downstream biochemical events. The range of biochemical functions of these proteins is correlated with differences in the way in which they respond to the binding of Ca2+. The EF-hand domains of calbindin D9k and calmodulin are homologous, yet they respond to the binding of calcium ions in a drastically different manner. A series of comparative analyses of their structures enabled the development of hypotheses about which residues in these proteins control the calcium-induced changes in conformation. To test our understanding of the relationship between protein sequence and structure, we specifically designed the F36G mutation of the EF-hand protein calbindin D9k to alter the packing of helices I and II in the apoprotein. The three-dimensional structure of apo F36G was determined in solution by nuclear magnetic resonance spectroscopy and showed that the design was successful. Surprisingly, significant structural perturbations also were found to extend far from the site of mutation. The observation of such long-range effects provides clear evidence that four-helix EF-hand domains should be treated as a single globally cooperative unit. A hypothetical mechanism for how the long-range effects are transmitted is described. Our results support the concept of energetic and structural coupling of the key residues that are crucial for a protein's fold and function.
Keywords: Calcium-binding protein; conformational change; EF-hand; mutagenesis; NMR spectroscopy; protein engineering
Abbreviations: CaBP, Ca2+-binding protein • CaM, calmodulin • calbindin, calbindin D9k • NMR, nuclear magnetic resonance • CSI, chemical shift index • NOE, nuclear Overhauser effect
CiteULike 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  N. Juranic, E. Atanasova, J. H. Streiff, S. Macura, and F. G. Prendergast Solvent-induced differentiation of protein backbone hydrogen bonds in calmodulin Protein Sci., July 1, 2007; 16(7): 1329 - 1337. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. M. Lakowski, G. M. Lee, M. Okon, R. E. Reid, and L. P. McIntosh Calcium-induced folding of a fragment of calmodulin composed of EF-hands 2 and 3 Protein Sci., June 1, 2007; 16(6): 1119 - 1132. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. J. Chazin The Impact of X-ray Crystallography and NMR on Intracellular Calcium Signal Transduction by EF-Hand Proteins: Crossing the Threshold from Structure to Biology and Medicine Sci. Signal., May 29, 2007; 2007(388): pe27 - pe27. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Sivaraja, T. K. S. Kumar, D. Rajalingam, I. Graziani, I. Prudovsky, and C. Yu Copper Binding Affinity of S100A13, a Key Component of the FGF-1 Nonclassical Copper-Dependent Release Complex Biophys. J., September 1, 2006; 91(5): 1832 - 1843. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. H. Sheehan, C. G. Bunick, H. Hu, P. A. Fagan, S. M. Meyn, and W. J. Chazin Structure of the N-terminal Calcium Sensor Domain of Centrin Reveals the Biochemical Basis for Domain-specific Function J. Biol. Chem., February 3, 2006; 281(5): 2876 - 2881. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. S. Yousef, W. A. Baase, and B. W. Matthews Use of sequence duplication to engineer a ligand-triggered, long-distance molecular switch in T4 lysozyme PNAS, August 10, 2004; 101(32): 11583 - 11586. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. A. Likic, E. E. Strehler, and P. R. Gooley Dynamics of Ca2+-saturated calmodulin D129N mutant studied by multiple molecular dynamics simulations Protein Sci., October 1, 2003; 12(10): 2215 - 2229. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
