Luminescent and substrate binding activities of firefly luciferase N-terminal domain

Tamotsu Zako, Keiichi Ayabe, Takahide Aburatani, Noriho Kamiya, Atsushi Kitayama, Hiroshi Ueda, Teruyuki Nagamune

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Firefly luciferase catalyzes highly efficient emission of light from the substrates luciferin, Mg-ATP, and oxygen. A number of amino acid residues are identified to be important for the luminescent activity, and almost all the key residues are thought to be located in the N-terminal domain (1-437), except one in the C-terminal domain, Lys529, which is thought to be critical for efficient substrate orientation. Here we show that the purified N-terminal domain still binds to the substrates luciferin and ATP with reduced affinity, and retains luminescent activity of up to 0.03% of the wild-type enzyme (WT), indicating that all the essential residues for the activity are located in the N-terminal domain. Also found is low luminescence enhancement by coenzyme A (CoA), which implies a lower product inhibition than in the WT enzyme. These findings have interesting implications for the light emission reaction mechanism of the enzyme, such as reaction intermediates, product inhibition, and the role of the C-terminal domain.

Original languageEnglish
Pages (from-to)183-189
Number of pages7
JournalBiochimica et Biophysica Acta - Proteins and Proteomics
Volume1649
Issue number2
DOIs
Publication statusPublished - Jul 30 2003

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Firefly Luciferases
Substrates
Enzymes
Adenosine Triphosphate
Light
Reaction intermediates
Light emission
Coenzyme A
Luminescence
Oxygen
Amino Acids

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biophysics
  • Biochemistry
  • Molecular Biology

Cite this

Luminescent and substrate binding activities of firefly luciferase N-terminal domain. / Zako, Tamotsu; Ayabe, Keiichi; Aburatani, Takahide; Kamiya, Noriho; Kitayama, Atsushi; Ueda, Hiroshi; Nagamune, Teruyuki.

In: Biochimica et Biophysica Acta - Proteins and Proteomics, Vol. 1649, No. 2, 30.07.2003, p. 183-189.

Research output: Contribution to journalArticle

Zako, Tamotsu ; Ayabe, Keiichi ; Aburatani, Takahide ; Kamiya, Noriho ; Kitayama, Atsushi ; Ueda, Hiroshi ; Nagamune, Teruyuki. / Luminescent and substrate binding activities of firefly luciferase N-terminal domain. In: Biochimica et Biophysica Acta - Proteins and Proteomics. 2003 ; Vol. 1649, No. 2. pp. 183-189.
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AU - Ueda, Hiroshi

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