Fluorescence properties of porcine odorant binding protein Trp 16 residue

Energy Transfer Studies between Trp Residues of Three Lipocalin Proteins Family, α1-Acid Glycoprotein, (Orosomucoid), β-Lactoglobulin and Porcine Odorant Binding Protein and the Fluorescent Probe, 1-Aminoanthracene (1-AMA)

“…The constant rate of the energy transfer (k t ) can be calculated from Eq. 5: where τ o is the mean fluorescence lifetime (= 4.778 ns for OBP [37] and 2.285 ns for α 1 -acid glycoprotein [38]. Equation 5 yields a value for k t equal to 0.4266×10 9 s −1 for OBP and 0.19×10 9 s −1 for α 1 -acid glycoprotein.…”

Novel biosensor system model based on fluorescence quenching by a fluorescent streptavidin and carbazole-labeled biotin

“…The DNAs of wild type and Tyr54, Tyr83, and Trp120 mutant streptavidins (Albani, ), DNAs of the tRNA that have a four‐base anticodon (Cornish et al ., ) and BFLAF‐pdCpA (Hohsaka et al ., ), were prepared as described previously. Alkaline phosphatase‐labeled anti‐mouse IgG and MagneHis™ Ni‐Particles were purchased from Promega.…”

“…In contrast, the unnatural amino acid mutagenesis method using amber and four‐base codons is an excellent method which can achieve site‐directed introduction of the functional unnatural amino acid that possessed fluorescent or redox‐active properties to the designed protein to provide a new function for the molecular biosensor (Cavalcanti et al ., ; Fan et al ., ; Witzens and Hochberg, ). In the four‐base codon method (Cornish et al ., ; Hohsaka et al ., ; Murakami et al ., ; Hohsaka and Sisido, ; Beene et al ., ; Saxl et al ., ; Albani, ), one or multiple four‐base codons were introduced into an assigned position of the target protein gene. The full‐length protein containing the unnatural amino acid will be produced when the four‐base codon is successfully decoded by the unnatural aminoacyl‐tRNA having the corresponding four‐base anticodon, but when the first three bases of the four‐base codon are decoded as a three‐base codon by a cognate naturally occurring aminoacyl‐tRNA, a frame shift occurs to cause the emergence of a stop codon, resulting in the termination of peptide elongation.…”

A novel molecular biosensor system model designed by using a couple composed of a fluorescent mutant binding protein and fluorophore-labeled ligand analogue based on the FRET mechanism

“…Molecular biosensors are being developed as suitable tools for different applications, including bioprocess control, food quality control, agriculture, the environment, military and in particular, for medical applications. [1][2][3][4][5] Functional unnatural proteins, which are proteins incorporating single or multiple functional unnatural amino acids, are applied to the biosensor and protein structural and functional analysis. Chemical modication is a general method for providing new functions such as uorescence, oxidationreduction and so forth to biological components.…”