Purification of the yellow fluorescent protein from vibrio fischeri and identity of the flavin chromophore

• Published in: Biochemical and biophysical research communications Vol. 146; no. 1; pp. 101 - 106
• Main Authors: Macheroux, P., Schmidt, K.U., Steinerstauch, P., Ghisla, S., Colepicolo, P., Buntic, R., Hastings, J.W.
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 15.07.1987; Elsevier
• Subjects: Applied sciences; chromophores; Exact sciences and technology; flavin; Flavin Mononucleotide; Fluorescence; Molecular Weight; Other techniques and industries; Spectrometry, Fluorescence; Vibrio - analysis; Vibrio fischeri; Viral Proteins - isolation & purification
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/0006-291X(87)90696-6

A low molecular weight protein (∼ 25,000 D) exhibiting a yellow fluorescence emission peaking at ∼ 540 nm was isolated from Vibrio fischeri (strain Y-1) and purified to apparent homogeneity. FMN is the chromophore, but it exhibits marked red shifts in both the absorption (λ max = 380, 460 nm) and the fluorescence emission. When added to purified luciferase from the same strain, which itself catalyzes an emission of blue-green light (λ max ∼ 495 nm), this protein induces a bright yellow luminescence (λ max ∼ 540 nm); this corresponds to the emission of the Y-1 strain in vivo . This yellow bioluminescence emission is thus ascribed to the interaction of these two proteins, and to the excitation of the singlet FMN bound to this fluorescent protein.