title: Relationship between stability and bioluminescence color of firefly luciferase.
abstract: Firefly luciferase catalyzes the oxidation of luciferin in the presence of ATP, Mg(2+) and molecular oxygen. The bioluminescence color of firefly luciferases is identified by the luciferase structure and assay conditions. Amongst different types of beetles, luciferase from Phrixotrix railroad worm (PhRE) with a unique additional residue (Arg353) naturally emits red bioluminescence color. By insertion of Arg356 in luciferase of Lampyris turkestanicus, corresponding to Arg353 in Phrixotrix hirtus, the color of the emitted light was changed to red. To understand the effect of this position on the bioluminescence color shift, four residues with similar sizes but different charges (Arg, Lys, Glu, and Gln) were inserted into Photinus pyralis luciferase. Comparison of mutants with native luciferase shows that mutation brought an increase in the content of secondary structure and globular compactness of (P. pylalis) luciferase. Comparative study of chemical denaturation of native and mutant luciferases by activity measurement, intrinsic and extrinsic fluorescence, circular dichroism, and DSC techniques revealed that insertion of positively charged residues (Arg, Lys) in the flexible loop (352-358) plays a significant role on the stability of (P. pyralis) luciferase and changes the light color to red.
authors.affiliation: Department of Biochemistry and Biophysics, Faculty of Biological Sciences, Tarbiat Modares University, P.O. Box 14115-175, Tehran, Iran.
authors.forename: Parvaneh
authors.initials: P
authors.lastname: Maghami
authors.forename: Bijan
authors.initials: B
authors.lastname: Ranjbar
authors.forename: Saman
authors.initials: S
authors.lastname: Hosseinkhani
authors.forename: Atiyeh
authors.initials: A
authors.lastname: Ghasemi
authors.forename: Ali
authors.initials: A
authors.lastname: Moradi
authors.forename: Pooria
authors.initials: P
authors.lastname: Gill
mesh_terms: D000818:Animals; D002942:Circular Dichroism; D048888:Fireflies / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D049409:Luciferases, Firefly / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D049449:Luminescence; D009154:Mutation; D011489:Protein Denaturation; D055550:Protein Stability; D014508:Urea / Q000378:metabolism
chemical_list: D014508:Urea; D049409:Luciferases, Firefly