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Replacements of Pro86 in phage T4 lysozyme extend an alpha-helix but do not alter protein stability.

Alber T, Bell JA, Sun DP, Nicholson H, Wozniak JA, Cook S, Matthews BW
Science 239 631-5 (1988)

Cited: 63 times
EuropePMC logo PMID: 3277275

Abstract

To investigate the relation between protein stability and the predicted stabilities of individual secondary structural elements, residue Pro86 in an alpha-helix in phage T4 lysozyme was replaced by ten different amino acids. The x-ray crystal structures of seven of the mutant lysozymes were determined at high resolution. In each case, replacement of the proline resulted in the formation of an extended alpha-helix. This involves a large conformational change in residues 81 to 83 and smaller shifts that extend 20 angstroms across the protein surface. Unexpectedly, all ten amino acid substitutions marginally reduce protein thermostability. This insensitivity of stability to the amino acid at position 86 is not simply explained by statistical and thermodynamic criteria for helical propensity. The observed conformational changes illustrate a general mechanism by which proteins can tolerate mutations.

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Related citations provided by authors (19)

  1. Structural studies of mutants of T4 lysozyme that alter hydrophobic stabilization.. Matsumura M, Wozniak JA, Sun DP, Matthews BW J Biol Chem 264 16059-66 (1989)
  2. High-Resolution Structure of the Temperature-Sensitive Mutant of Phage Lysozyme, Arg 96 (Right Arrow) His. Weaver LH, Gray TM, Gruetter MG, Anderson DE, Wozniak JA, Dahlquist FW, Matthews BW Biochemistry 28 3793- (1989)
  3. Contributions of left-handed helical residues to the structure and stability of bacteriophage T4 lysozyme.. Nicholson H, Söderlind E, Tronrud DE, Matthews BW J Mol Biol 210 181-93 (1989)
  4. Hydrophobic stabilization in T4 lysozyme determined directly by multiple substitutions of Ile 3.. Matsumura M, Becktel WJ, Matthews BW Nature 334 406-10 (1988)
  5. Enhanced protein thermostability from designed mutations that interact with alpha-helix dipoles.. Nicholson H, Becktel WJ, Matthews BW Nature 336 651-6 (1988)
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  11. Temperature-sensitive mutations of bacteriophage T4 lysozyme occur at sites with low mobility and low solvent accessibility in the folded protein.. Alber T, Sun DP, Nye JA, Muchmore DC, Matthews BW Biochemistry 26 3754-8 (1987)
  12. Common precursor of lysozymes of hen egg-white and bacteriophage T4.. Matthews BW, Grütter MG, Anderson WF, Remington SJ Nature 290 334-5 (1981)
  13. Crystallographic Determination of the Mode of Binding of Oligosaccharides to T4 Bacteriophage Lysozyme. Implications for the Mechanism of Catalysis. Anderson WF, Gruetter MG, Remington SJ, Weaver LH, Matthews BW J. Mol. Biol. 147 523- (1981)
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  17. Comparison of the predicted and observed secondary structure of T4 phage lysozyme.. Matthews BW Biochim Biophys Acta 405 442-51 (1975)
  18. The three dimensional structure of the lysozyme from bacteriophage T4.. Matthews BW, Remington SJ Proc Natl Acad Sci U S A 71 4178-82 (1974)
  19. Crystallographic Data for Lysozyme from Bacteriophage T4. Matthews BW, Dahlquist FW, Maynard AY J. Mol. Biol. 78 575- (1973)

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