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

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• 作者：Hisakage Funabashi (1) (3)
  Satoshi Yanagi (1)
  Shigeya Suzuki (2)
  Masayasu Mie (1) (4)
  Eiry Kobatake (1) (4)
  
  1. Department of Biological Information ; Graduate School of Bioscience and Biotechnology ; Tokyo Institute of Technology ; 4259 Nagatsuta ; Midori-ku ; Yokohama ; 226-8501 ; Japan
  3. Institute for Sustainable Sciences and Development ; Hiroshima University ; 2-313 ; Kagamiyama ; Higashihiroshima ; Hiroshima ; 739-8527 ; Japan
  2. Biochemical Department ; Kikkoman Food Products Company ; 399 Noda ; Noda-shi ; Chiba ; 278-0037 ; Japan
  4. Department of Environmental Chemistry and Engineering ; Graduate School of Interdisciplinary Science and Engineering ; Tokyo Institute of Technology ; 4259 Nagatsuta ; Midori-ku ; Yokohama ; 226-8501 ; Japan
  
• 关键词：DNA scaffolds ; Enzyme assembly ; Firefly luciferase ; Proximity effect ; Pyruvate phosphate dikinase ; Zinc finger motifs
• 刊名：Biotechnology Letters
• 出版年：2015
• 出版时间：January 2015
• 年：2015
• 卷：37
• 期：1
• 页码：109-114
• 全文大小：368 KB
• 参考文献：1. Beerli RR, Barbas CF III (2002) Engineering polydactyl zinc-finger transcription factors. Nat Biotechnol 20:135鈥?41 CrossRef
  2. Blancafort P, Segal DJ, Barbas CF III (2004) Designing transcription factor architectures for drug discovery. Mol Pharmacol 66:1361鈥?371 CrossRef
  3. Ebihara T, Takayama H, Yanagida Y, Kobatake E, Aizawa M (2002) Thermostabilization of protein A-luciferase fusion protein by single amino acid mutation. Biotechnol Lett 24:147鈥?49 CrossRef
  4. Eisaki N, Tatsumi H, Murakami S, Horiuchi T (1999) Pyruvate phosphate dikinase from a thermophilic actinomyces / Microbispora rosea subsp. / aerata: purification, characterization and molecular cloning of the gene. Biochim Biophys Acta 1431:363鈥?73 CrossRef
  5. Fu J, Liu M, Liu Y, Woodbury NW, Yan H (2012a) Interenzyme substrate diffusion for an enzyme cascade organized on spatially addressable DNA nanostructures. J Am Chem Soc 134:5516鈥?519 CrossRef
  6. Fu J, Liu M, Liu Y, Yan H (2012b) Spatially-interactive biomolecular networks organized by nucleic acid nanostructures. Acc Chem Res 45:1215鈥?226 CrossRef
  7. M眉ller J, Niemeyer CM (2008) DNA-directed assembly of artificial multienzyme complexes. Biochem Biophys Res Commun 377:62鈥?7 CrossRef
  8. Negi S, Imanishi M, Matsumoto M, Sugiura Y (2008) New redesigned zinc-finger proteins: design strategy and its application. Chemistry 14:3236鈥?249 CrossRef
  9. Niemeyer CM (2010) Semisynthetic DNA鈥損rotein conjugates for biosensing and nanofabrication. Angew Chem Int Ed Engl 49:1200鈥?216 CrossRef
  10. Niemeyer CM, Koehler J, Wuerdemann C (2002) DNA-directed assembly of bienzymic complexes from in vivo biotinylated NAD(P)H:FMN oxidoreductase and luciferase. ChemBioChem 3:242鈥?45 CrossRef
  11. Ooi AT, Stains CI, Ghosh I, Segal DJ (2006) Sequence-enabled reassembly of beta-lactamase (SEER-LAC): a sensitive method for the detection of double-stranded DNA. Biochemistry 45:3620鈥?625 CrossRef
  12. Pavletich NP, Pabo CO (1991) Zinc finger-DNA recognition: crystal structure of a Zif268鈥揇NA complex at 2.1 脜. Science 252:809鈥?17 CrossRef
  13. Wilner OI, Shimron S, Weizmann Y, Wang ZG, Willner I (2009a) Self-assembly of enzymes on DNA scaffolds: en route to biocatalytic cascades and the synthesis of metallic nanowires. Nano Lett 9:2040鈥?043 CrossRef
  14. Wilner OI, Weizmann Y, Gill R, Lioubashevski O, Freeman R, Willner I (2009b) Enzyme cascades activated on topologically programmed DNA scaffolds. Nat Nanotechnol 4:249鈥?54 CrossRef
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Microbiology
  Biotechnology
  Applied Microbiology
  Biochemistry
  
• 出版者：Springer Netherlands
• ISSN：1573-6776

文摘

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.