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Your search keyword '"Mie, Masayasu"' showing total 22 results
Start Over Author "Mie, Masayasu" Journal biotechnology letters
22 results on '"Mie, Masayasu"'

19. Assembly of zinc finger motif-fused enzymes on a dsDNA scaffold for catalyzing consecutive reactions with a proximity effect

Author

Funabashi, Hisakage, Yanagi, Satoshi, Suzuki, Shigeya, mie, masayasu, and KOBATAKE, EIRY

Subjects
chemistry.chemical_classification, Zinc finger, Recombinant Fusion Proteins, Bioengineering, Zinc Fingers, General Medicine, Protein engineering, DNA, Protein Engineering, Applied Microbiology and Biotechnology, Zinc finger nuclease, Pyruvate, Orthophosphate Dikinase, Pyruvate, phosphate dikinase, chemistry.chemical_compound, Enzyme, Adenosine Triphosphate, Biochemistry, chemistry, Luciferases, Firefly, Luciferase, Binding site, Biotechnology
Abstract

The feasibility of assembling enzymes, catalyzing consecutive reactions, on to a double-stranded DNA (dsDNA) scaffold utilizing zinc finger motifs is described. The catalytic activities of two zinc finger motif-fused enzymes catalyzing a bioluminescence reaction with energy recycling, namely pyruvate phosphate dikinase and firefly luciferase, have been evaluated. Bioluminescence measurements with dsDNA scaffolds coding a different distance between the binding sites for each zinc finger motif-fused enzyme confirmed the effect of the distance, proving the proximity effect of ATP recycling presumed to be the result of efficient intermediate diffusion. Thus, fusion to zinc finger motifs offers a promising option for the assembly of bi-enzymes, catalyzing a consecutive reaction, onto a dsDNA scaffold with a proximity effect.

Published
2014

20. Evaluation of small ligand-protein interaction by ligation reaction with DNA-modified ligand

Author

Sugita, R., mie, masayasu, Funabashi, H., and KOBATAKE, EIRY

Subjects
Streptavidin, DNA Ligases, Biotin, Bioengineering, Applied Microbiology and Biotechnology, Models, Biological, Polymerase Chain Reaction, chemistry.chemical_compound, A-DNA, chemistry.chemical_classification, DNA ligase, Hybridization probe, Proteins, General Medicine, DNA, chemistry, Biochemistry, Biophysics, Ligation, Molecular probe, DNA Probes, Digoxigenin, Biotechnology, Protein Binding
Abstract

A method for the evaluation of interactions between protein and ligand using DNA-modified ligands, including signal enhancement of the DNA ligation reactions, is described. For proof of principle, a DNA probe modified by biotin was used. Two DNA probes were prepared with complementary sticky-ends. While one DNA probe was modified at the 5′-end of the sticky-end, the other was not modified. The probes could be ligated together by T4 DNA ligase along the strand without biotin modification. However, in the presence of streptavidin or anti-biotin Fab, the ligation reaction joining the two probes could not occur on either strand.

Published
2009

22. Functional decoration of elastin-like polypeptides-based nanoparticles with a modular assembly via isopeptide bond formation.

Author

Yamaguchi J, Nishida K, Kobatake E, and Mie M

Subjects
Green Fluorescent Proteins chemistry, Green Fluorescent Proteins genetics, Green Fluorescent Proteins metabolism, Temperature, Recombinant Fusion Proteins chemistry, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins metabolism, Elastin-Like Polypeptides, Elastin chemistry, Elastin metabolism, Nanoparticles chemistry, Peptides chemistry, Peptides metabolism
Abstract

Temperature-responsive elastin-like polypeptides (ELPs) exhibit a low critical solution temperature-type phase transition and offer potential as useful materials for the construction of nanoparticles. Herein, we developed a novel decoration method for ELP-based nanoparticles via isopeptide bond formation with the SnoopTag/SnoopCatcher system that is not affected by the heating process required for particle formation. A mixture of a fusion protein of ELP and poly(aspartic acid) (poly(D)), known as ELP-poly(D), and ELP-poly(D) fused with SnoopCatcher (ELP-poly(D)-SnC) formed protein nanoparticles as a result of the temperature responsiveness of ELP, with the resultant nanoparticles displaying the SnoopCatcher binding domain on their surfaces. In the present study, two model proteins fused to SnoopTag were displayed on the surfaces of protein nanoparticles constructed from ELP-poly(D)-SnC and ELP-poly(D). The model proteins are enhanced green fluorescent protein (EGFP) and Renilla luciferace (Rluc), which exhibits luminescent capability and weak thermostability, respectively. EGFP on the particle surface was found to retain 48.7% activity, while Rluc exhibited almost full activity, as calculated from the binding efficiency and nanoparticle activities recovered after purification. ELP-based nanoparticles containing the SnoopTag/SnoopCatcher system offer the opportunity for particle decoration with a wide range of functional proteins via isopeptide bond formation., Competing Interests: Declarations. Competing interests: The authors have no relevant financial or non-financial interests to disclose., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Published
2024
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