Abstract

The principal objective of this proposal is to use T4 lysozyme as a model system to better understand the factors that determine the folding, stability, structure and function of proteins. The specific research to be accomplished includes the following: (a) An attempt will be made to simplify the protein folding problem by identifying which residues, or combinations of residues, in T4 lysozyme are critical for folding and stability. We want to understand not only how given residues contribute to stability, but also the signals, if any, that define the elements of secondary structure. Ultimately we would like to reduce the amino acid sequence of T4 lysozyme to the simplest form that will still give a folded, functional protein. (b) Methionine substitution, together with other nonpolar replacements, will be used to better understand the core-packing interactions that are critical to protein folding. (c) Methods will be developed and tested to improve protein stability. (d) Cavities within T4 lysozyme will be exploited both to understand protein-ligand interaction and to engineer novel active sites. (e) The role of strain within the protein will be systematically analyzed.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM021967-23
Application #
2391832
Study Section
Molecular and Cellular Biophysics Study Section (BBCA)
Project Start
1978-06-01
Project End
2000-03-31
Budget Start
1997-04-01
Budget End
1998-03-31
Support Year
23
Fiscal Year
1997
Total Cost
Indirect Cost

  Institution

Name
University of Oregon
Department
Type
Organized Research Units
DUNS #
948117312
City
Eugene
State
OR
Country
United States
Zip Code
97403

  Publications

Baase, Walter A; Liu, Lijun; Tronrud, Dale E et al. (2010) Lessons from the lysozyme of phage T4. Protein Sci 19:631-41
Liu, Lijun; Marwitz, Adam J V; Matthews, Brian W et al. (2009) Boron mimetics: 1,2-dihydro-1,2-azaborines bind inside a nonpolar cavity of T4 lysozyme. Angew Chem Int Ed Engl 48:6817-9
Mooers, Blaine H M; Tronrud, Dale E; Matthews, Brian W (2009) Evaluation at atomic resolution of the role of strain in destabilizing the temperature-sensitive T4 lysozyme mutant Arg 96 --> His. Protein Sci 18:863-70
Liu, Lijun; Baase, Walter A; Matthews, Brian W (2009) Halogenated benzenes bound within a non-polar cavity in T4 lysozyme provide examples of I...S and I...Se halogen-bonding. J Mol Biol 385:595-605
Liu, Lijun; Baase, Walter A; Michael, Miya M et al. (2009) Use of stabilizing mutations to engineer a charged group within a ligand-binding hydrophobic cavity in T4 lysozyme. Biochemistry 48:8842-51
Matthews, Brian W; Liu, Lijun (2009) A review about nothing: are apolar cavities in proteins really empty? Protein Sci 18:494-502
Mooers, Blaine H M; Baase, Walter A; Wray, Jonathan W et al. (2009) Contributions of all 20 amino acids at site 96 to the stability and structure of T4 lysozyme. Protein Sci 18:871-80
Liu, Lijun; Quillin, Michael L; Matthews, Brian W (2008) Use of experimental crystallographic phases to examine the hydration of polar and nonpolar cavities in T4 lysozyme. Proc Natl Acad Sci U S A 105:14406-11
Collins, Marcus D; Quillin, Michael L; Hummer, Gerhard et al. (2007) Structural rigidity of a large cavity-containing protein revealed by high-pressure crystallography. J Mol Biol 367:752-63
Quillin, Michael L; Wingfield, Paul T; Matthews, Brian W (2006) Determination of solvent content in cavities in IL-1beta using experimentally phased electron density. Proc Natl Acad Sci U S A 103:19749-53

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