The lysozyme from bacteriophage T4 will be used as a model system to understand the factors that determine the folding, stability, activity and three-dimensional structures of proteins. The specific research to be accomplished is as follows: 1. Mutant lysozymes with altered stability (and/or catalytic activity) will be selected and studied in detail. These mutant lysozymes will include not only temperature-sensitive lysozymes but also lysozymes that are more thermally stable than the wild-type enzyme. The three-dimensional structures of these mutant lysozymes will be determined and compared with the wild-type lysozyme structure. Changes in structure will be correlated with changes in stability and activity. 2. Oligonucleotide-directed mutagenesis will be used to introduce designed alterations in the lysozyme structure. By making a series of selected amino acid replacements at one site we will discriminate between the contributions of different interactions at that site. Site-directed mutagenesis will also be used to test the importance of factors such as hydrogen bonding, secondary structure stabilization, hydrophobic interactions, and steric hindrance in protein stabilization. 3. The evolution of lysozyme structure and function will be investigated by determining and comparing the three-dimensional structures of lysozymes that have dissimilar amino acid sequences. In particular, we will continue structural studies of goose egg-white lysozyme and its relationship to chicken and phage lysozymes.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
5R01GM021967-14
Application #
3270845
Study Section
Biophysics and Biophysical Chemistry B Study Section (BBCB)
Project Start
Project End
Budget Start
Budget End
Support Year
14
Fiscal Year
1988
Total Cost
Indirect Cost
Name
University of Oregon
Department
Type
Organized Research Units
DUNS #
948117312
City
Eugene
State
OR
Country
United States
Zip Code
97403
Baase, Walter A; Liu, Lijun; Tronrud, Dale E et al. (2010) Lessons from the lysozyme of phage T4. Protein Sci 19:631-41
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
Matthews, Brian W; Liu, Lijun (2009) A review about nothing: are apolar cavities in proteins really empty? Protein Sci 18:494-502
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
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
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
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; 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

Showing the most recent 10 out of 112 publications