Structure of flavoprotein FP390 from a luminescent bacterium Photobacterium phosphoreum refined at 2.7 Å resolution

Kita, Akiko; Kasai, Shinji; Miyata, Masayuki; Miki, Kazuhiko

doi:10.1107/s0907444995009796

Cited by 10 publications

(14 citation statements)

References 23 publications

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“…The first two bands suggested that the NdeI and/or BamHI sites existed in the PCR product. Nucleotide sequencing showed that the coding region of the cloned PCR product consisted of 696 nucleotides including the stop codon, in exact agreement with the reported size of the luxF gene (10,18). Figure 1 shows the alignment of the amino acid sequence of bmFP deduced from the sequenced gene with that for the luxF gene product (18).…”

Section: Gene Cloningsupporting

confidence: 77%

“…The ratio of A 268 to A 386 for bmFP is 3.87, close to the values of about 3.8 for NFP (7) and 3.9 for FP 390 (8). Based on these values, it is expected that one bmFP molecule binds four FMN-myristate molecules, as do NFP and FP 390 (16,18).…”

Section: Absorption and Fluorescence Properties Of Bmfpsupporting

confidence: 69%

“…The 19 N-terminal amino acids of purified bmFP were determined to be MNKWNYGVFFVNFYNKGQQ. This sequence corresponds exactly to that of the luxF gene product of P. phosphoreum (NCMB844) (10,11) and P. phosphoreum (IFO13896) (18). This implied that the apo-bmFP is homologous to the luxF gene product, even though the fluorescence properties of the holo-bmFP and the LuxF proteins, such as NFP and FP 390 , are significantly different.…”

Section: Gene Cloningmentioning

confidence: 62%

“…The amino acid sequences of the LuxF and LuxN proteins, deduced on the basis of the sequences determined for the two genes, are very similar to each other. X-ray diffraction studies have revealed that NFP (14)(15)(16) and FP 390 (17,18) are homodimers, and both carry two FMN-myristate adducts per monomer. It has also been shown that the non-or weakly fluorescent properties are caused by stacking of fluorescence-quenching aromatic amino acid residues onto the chromophores (14,16).…”

Section: Introductionmentioning

confidence: 99%

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Properties of the Bimodal Fluorescent Protein Produced by Photobacterium phosphoreum

Karatani¹,

Konaka²,

Katsukawa³

2007

Photochemistry and Photobiology

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A fluorescent protein isolated from the deep‐sea luminous bacterium Photobacterium phosphoreum strain bmFP has been purified, cloned and sequenced. The protein is 96.5% identical in amino acid sequence to FP390, the weakly fluorescent flavoprotein encoded by the luxF gene characteristic of Photobacterium species. Similar to FP390, bmFP is a dimer of two homologous subunits binding four FMN‐myristate chromophores but has the distinctive feature of emitting a bimodal fluorescence with maxima at about 488 and 517 nm, hence the name bmFP. For both bands of this fluorescence, the excitation spectrum exhibits a peak at 336 nm, not corresponding to its flavin‐like absorption spectrum. Heating of bmFP in urea resulted in a decrease in the intensity of the 488 nm band along with the appearance of a new fluorescence peaking at 423 nm, partially reversible upon the removal of the urea. Upon complete denaturation, either by heat or guanidium chloride at 65°C, fluorescence characteristic of both free flavin and this 423 nm species appears. It is speculated that chromophores in different states of protonation, associated with a single protein, are responsible for the unusual spectral properties of bmFP.

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