X-ray structural and simulation analysis of a protein mutant: The value of a combined approach
Corresponding Author
Carla Mattos
Department of Molecular and Structural Biochemistry, North Carolina State University, Raleigh, North Carolina
Carla Mattos, Department of Molecular and Structural Biochemistry, North Carolina State University, 128 Polk Hall, Raleigh, NC 27695-7622===
Martin Karplus, Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford St., Cambridge, MA 02138===
Search for more papers by this authorJustin D. Cohen
Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts
Search for more papers by this authorDavid F. Green
Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts
Search for more papers by this authorBruce Tidor
Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts
Search for more papers by this authorCorresponding Author
Martin Karplus
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts
Laboratoire de Chimie Biophysique, Institut le Bel Universite Louis Pasteur, Strasbourg, France
Carla Mattos, Department of Molecular and Structural Biochemistry, North Carolina State University, 128 Polk Hall, Raleigh, NC 27695-7622===
Martin Karplus, Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford St., Cambridge, MA 02138===
Search for more papers by this authorCorresponding Author
Carla Mattos
Department of Molecular and Structural Biochemistry, North Carolina State University, Raleigh, North Carolina
Carla Mattos, Department of Molecular and Structural Biochemistry, North Carolina State University, 128 Polk Hall, Raleigh, NC 27695-7622===
Martin Karplus, Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford St., Cambridge, MA 02138===
Search for more papers by this authorJustin D. Cohen
Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts
Search for more papers by this authorDavid F. Green
Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts
Search for more papers by this authorBruce Tidor
Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts
Search for more papers by this authorCorresponding Author
Martin Karplus
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts
Laboratoire de Chimie Biophysique, Institut le Bel Universite Louis Pasteur, Strasbourg, France
Carla Mattos, Department of Molecular and Structural Biochemistry, North Carolina State University, 128 Polk Hall, Raleigh, NC 27695-7622===
Martin Karplus, Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford St., Cambridge, MA 02138===
Search for more papers by this authorAbstract
The effect of the mutation Arg 96 to His on the stability of bacteriophage T4 lysozyme has been previously studied by calorimetric experiments, X-ray crystallography, and free energy simulation techniques. The experimental and calculated values for the difference between the free energy of denaturation of the mutant and the wild type are in reasonable agreement. However, the two approaches led to different explanations for the loss in stability. To analyze the differences, a series of refinements based on the crystallographic data were performed, a number of aspects of the simulations were reexamined, and continuum electrostatic calculations were done to complement the latter. The results of those comparisons provide a better understanding of the origin of the free energy difference in this mutant. Furthermore, they show the importance of the combined use of simulations and crystallography for interpreting the effects of mutations on the energetics of the system. Proteins 2004. © 2004 Wiley-Liss, Inc.
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