Stabilization of lysozyme by the introduction of gly-pro sequence

Tadashi Ueda, Tomohiro Tamura, Yoshitake Maeda, Yoshio Hashimoto, Takeyoshi Miki, Hidenori Yamada, Taiji Imoto

Research output: Contribution to journalArticle

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Abstract

Three mutant lysozymes where the Asp101 - Gly102 sequence of lysozyme was converted to Asp101-Pro102, Gly101-Pro102 and Pro101-Gly102 were prepared to investigate the effect of proline residues on the stabilization of proteins. The free energy changes of lysozymes for the unfolding in aqueous solution at pH 5.5 and 35°C were 10.0, 10.1, 11.0 and 7.7 kcal/mol for wild type, Asp101Pro102, Gly101Pro102 and Pro101Gly102 lysozyme respectively. When the energy level in the unfolded state of wild type lysozyme was fixed at a standard level, the energy levels in the folded state of Asp101Pro102 and Pro101Gly102 lysozymes were found to be higher than that of wild type lysozyme on the basis of ΔGD(H2O) and entropy losses of their polypeptide chains in the unfolded state. The presence of some strain in the folded state of these lysozymes was supported by both the calculation of conformational energy for a trans-L-prolyl residue [Schimmel, P.R. and Flory,P.J. (1968) J. Mol. Biol, 34, 105 -120] and the analysis of structures of energy-minimized mutant lysozymes. Therefore, it is concluded that the formation of the Gly-Pro sequence is effective in avoiding possible strain in the folded state of a protein caused by the introduction of proline residue(s).

Original languageEnglish
Pages (from-to)183-187
Number of pages5
JournalProtein Engineering, Design and Selection
Volume6
Issue number2
DOIs
Publication statusPublished - Feb 1 1993

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glycylproline
Muramidase
Enzymes
Stabilization
Proline
Electron energy levels
Proteins
Polypeptides
Entropy
Free energy

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Molecular Biology

Cite this

Stabilization of lysozyme by the introduction of gly-pro sequence. / Ueda, Tadashi; Tamura, Tomohiro; Maeda, Yoshitake; Hashimoto, Yoshio; Miki, Takeyoshi; Yamada, Hidenori; Imoto, Taiji.

In: Protein Engineering, Design and Selection, Vol. 6, No. 2, 01.02.1993, p. 183-187.

Research output: Contribution to journalArticle

Ueda, T, Tamura, T, Maeda, Y, Hashimoto, Y, Miki, T, Yamada, H & Imoto, T 1993, 'Stabilization of lysozyme by the introduction of gly-pro sequence', Protein Engineering, Design and Selection, vol. 6, no. 2, pp. 183-187. https://doi.org/10.1093/protein/6.2.183
Ueda, Tadashi ; Tamura, Tomohiro ; Maeda, Yoshitake ; Hashimoto, Yoshio ; Miki, Takeyoshi ; Yamada, Hidenori ; Imoto, Taiji. / Stabilization of lysozyme by the introduction of gly-pro sequence. In: Protein Engineering, Design and Selection. 1993 ; Vol. 6, No. 2. pp. 183-187.
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