Lysozyme requires fluctuation of the active site for the manifestation of activity

Taiji Imoto, Tadashi Ueda, Tomohiro Tamura, Yoshimasa Isakari, Yoshito Abe, Makoto Inoue, Takeyoshi Miki, Keiichi Kawano, Hidenori Yamada

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Mutations around His15 which lie far away from the active site, stimulated glycol chitin activity of lysozyme at physiological temperature. Del-Argl4Hisl5 lysozyme, a mutant lysozyme whose Argl4 and Hisl5 were deleted together, and has the highest activity among these mutant lysozymes, had a similar binding ability to a trimer of N-acetyl-glucosamine, a substrate analogue, relative to native lysozyme. This suggests that the increased activity was due to an increased kcat in the catalysis reaction. The H-D exchange rate of the N-1 proton in the Trp63 which is located in the active site cleft, was enhanced in the Del-Argl4Hisl5 lysozyme, while 2-D proton NMR analysis revealed no conformational change around Trp63. We conclude that some sort of fluctuation at the active site might be required for the manifestation of activity. This theory is supported by the finding that the Del-Argl4Hisl5 lysozyme showed a shift in temperature dependency of activity to lower temperatures compared with that of native lysozyme.

Original languageEnglish
Pages (from-to)743-748
Number of pages6
JournalProtein Engineering, Design and Selection
Volume7
Issue number6
DOIs
Publication statusPublished - Jun 1 1994

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Muramidase
Catalytic Domain
Enzymes
Temperature
Protons
Glucosamine
Chitin
Glycols
Catalysis
Ion exchange
Thermodynamic properties
Nuclear magnetic resonance
Hydrogen
Mutation
Substrates

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Molecular Biology

Cite this

Lysozyme requires fluctuation of the active site for the manifestation of activity. / Imoto, Taiji; Ueda, Tadashi; Tamura, Tomohiro; Isakari, Yoshimasa; Abe, Yoshito; Inoue, Makoto; Miki, Takeyoshi; Kawano, Keiichi; Yamada, Hidenori.

In: Protein Engineering, Design and Selection, Vol. 7, No. 6, 01.06.1994, p. 743-748.

Research output: Contribution to journalArticle

Imoto, Taiji ; Ueda, Tadashi ; Tamura, Tomohiro ; Isakari, Yoshimasa ; Abe, Yoshito ; Inoue, Makoto ; Miki, Takeyoshi ; Kawano, Keiichi ; Yamada, Hidenori. / Lysozyme requires fluctuation of the active site for the manifestation of activity. In: Protein Engineering, Design and Selection. 1994 ; Vol. 7, No. 6. pp. 743-748.
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AU - Miki, Takeyoshi

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