|
KMID : 0380220070400040452
|
|
Journal of Biochemistry and Molecular Biology
2007 Volume.40 No. 4 p.452 ~ p.458
|
|
|
Critical Role of Glu175 on Stability and Folding of Bacterial Luciferase:Stopped-flow Fluorescence Study
|
|
Shirazy Najmeh Hadizadeh
Ranjbar Bijan
Hosseinkhani Saman
Khalifeh Khosrow
Madvar Ali Riahi
Naderi-Manesh Hossein
|
|
|
Abstract
|
|
|
|
Bacterial luciferase is a heterodimeric enzyme, which catalyzes the light emission reaction, utilizing reduced FMN (FMNH2), a long chain aliphatic aldehyde and O2, 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 α 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.
|
|
|
KEYWORD
|
|
|
Anisotropy, Bacterial luciferase, Stopped-flow fluorescence, φ-value
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|
|