title: C6-Deoxy coelenterazine analogues as an efficient substrate for glow luminescence reaction of nanoKAZ: the mutated catalytic 19 kDa component of Oplophorus luciferase.
abstract: The codon-optimized gene for the mutated 19 kDa protein (nanoKAZ), which is the catalytic component of Oplophorus luciferase, was expressed in Escherichia coli cells and the recombinant protein was highly purified. The secretory expression of nanoKAZ from CHO-K1 cells was performed by fusing the secretory signal peptide sequence of Gaussia luciferase to the amino-terminus of nanoKAZ. The substrate specificity for the purified nanoKAZ and the nanoKAZ secreted into the cultured medium was determined, indicating that bis-coelenterazine (bis-CTZ) and newly synthesized 6h-f-coelenterazine (6h-f-CTZ) are an efficient substrate for the glow luminescence reaction of nanoKAZ.
authors.affiliation: Yokohama Research Center, JNC Co., 5-1 Okawa, Kanazawa-ku, Yokohama 236-8605, Japan. sinouye@jnc-corp.co.jp
authors.forename: Satoshi
authors.initials: S
authors.lastname: Inouye
authors.forename: Jun-ichi
authors.initials: J
authors.lastname: Sato
authors.forename: Yuiko
authors.initials: Y
authors.lastname: Sahara-Miura
authors.forename: Suguru
authors.initials: S
authors.lastname: Yoshida
authors.forename: Hajime
authors.initials: H
authors.lastname: Kurakata
authors.forename: Takamitsu
authors.initials: T
authors.lastname: Hosoya
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D020134:Catalytic Domain; D033342:Copepoda; D006224:Cricetinae; D003412:Cricetulus; D033364:Decapoda / Q000201:enzymology*; D004926:Escherichia coli / Q000378:metabolism; D007093:Imidazoles / Q000737:chemistry / Q000378:metabolism*; D008156:Luciferases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D049449:Luminescence; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D021382:Protein Sorting Signals; D011719:Pyrazines / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D007093:Imidazoles; D050505:Mutant Proteins; D021382:Protein Sorting Signals; D011719:Pyrazines; D011993:Recombinant Fusion Proteins; C017144:coelenterazine; D008156:Luciferases
keywords: 19kOLase; CTZ; Codon-optimized gene; Coelenterazine; FWHM; Furimazine; GLase; Gaussia; Gaussia luciferase; I(max); Reporter assay; coelenterazine; full width at half maximum; maximum intensity of luminescence; nanoKAZ; relative light units; rlu; the catalytic 19kDa protein of Oplophorus luciferase; the mutated 19kOLase