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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Critical Role of Glu175 on Stability and Folding of Bacterial Luciferase: Stopped-flow Fluorescence Study

  • Shirazy, Najmeh Hadizadeh (Department of Biophysics & Biochemistry, Faculty of sciences, Tarbiat Modares University) ;
  • Ranjbar, Bijan (Department of Biophysics & Biochemistry, Faculty of sciences, Tarbiat Modares University) ;
  • Hosseinkhani, Saman (Department of Biophysics & Biochemistry, Faculty of sciences, Tarbiat Modares University) ;
  • Khalifeh, Khosrow (Department of Biophysics & Biochemistry, Faculty of sciences, Tarbiat Modares University) ;
  • Madvar, Ali Riahi (Department of Biophysics & Biochemistry, Faculty of sciences, Tarbiat Modares University) ;
  • Naderi-Manesh, Hossein (Department of Biophysics & Biochemistry, Faculty of sciences, Tarbiat Modares University)
  • Published : 2007.07.31
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Abstract

Bacterial luciferase is a heterodimeric enzyme, which catalyzes the light emission reaction, utilizing reduced FMN (FMNH2), a long chain aliphatic aldehyde and $O_2$, to produce green-blue light. This enzyme can be readily classed as slow or fast decay based on their rate of luminescence decay in a single turnover. Mutation of Glu175 in $\alpha$ subunit to Gly converted slow decay Xenorhabdus Luminescence luciferase to fast decay one. The following studies revealed that changing the luciferase flexibility and lake of Glu-flavin interactions are responsible for the unusual kinetic properties of mutant enzyme. Optical and thermodynamics studies have caused a decrease in free energy and anisotropy of mutant enzyme. Moreover, the role of Glu175 in transition state of folding pathway by use of stopped-flow fluorescence technique has been studied which suggesting that Glu175 is not involved in transition state of folding and appears as surface residue of the nucleus or as a member of one of a few alternative folding nuclei. These results suggest that mutation of Glu175 to Gly extended the structure of Xenorhabdus Luminescence luciferase, locally.

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References

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