Abstract
We propose a novel method to prepare a DNA–protein conjugate using histidine-tag (His-tag) chemistry. Oligo-DNA was modified with nitrilotriacetate (NTA), which has high affinity to a His-tag on recombinant protein via the complexation of Ni2+. Investigations using a microplate which displayed a complementary DNA-strand revealed that a NTA-modified DNA–protein conjugate was formed and immobilized in the presence of Ni2+ on the microplate. We then adopted alkaline phosphatase (AP) as a model protein, and application of the DNA–AP conjugate was demonstrated in a thrombin aptamer-based detection system with a detection limit of approximately 10 nM.
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References
Beissenhirtz MK, Willner I (2006) DNA-based machines. Org Biomol Chem 4:3392–3401
Bock LC, Griffin LC, Latham JA, Vermaas EH, Toole JJ (1992) Selection of single-stranded DNA molecules that bind and inhibit human thrombin. Nature 355:564–566
Boozer C, Ladd J, Chen S, Yu Q, Homola J, Jiang S (2004) DNA directed protein immobilization on mixed ssDNA/oligo(ethylene glycol) self-assembled monolayers for sensitive biosensor. Anal Chem 76:6967–6972
Drolet DW, MoonMcDermott L, Romig TS (1996) An enzyme-linked oligonucleotide assay. Nat Biotechnol 14:1021–1025
Ellington AD, Szostak JW (1990) In vitro selection of RNA molecules that bind specific ligands. Nature 346:818–822
Hainfeld JF, Liu WQ, Halsey CMR, Freimuth P, Powell RD (1999) Ni–NTA–gold clusters target his-tagged proteins. J Struct Biol 127:185–198
Maruyama T, Hosogi T, Goto M (2007) Sequence-selective extraction of single-strand DNA using DNA-functionalized reverse micelles. Chem Commun 43:4450–4452
Niemeyer CM (2001) Bioorganic applications of semisynthetic DNA–protein conjugates. Chem Eur J 7:3189–3195
Niemeyer CM (2002) The developments of semisynthetic DNA–protein conjugates. Trends Biotechnol 20:395–401
Nutiu R, Li YF (2003) Structure-switching signaling aptamers. J Am Chem Soc 125:4771–4778
Saghatelian A, Guckian KM, Thayer DA, Ghadiri MR (2003) DNA detection and signal amplification via an engineered allosteric enzyme. J Am Chem Soc 125:344–345
Schmid EL, Keller TA, Dienes Z, Vogel H (1997) Reversible oriented surface immobilization of functional proteins on oxide surfaces. Anal Chem 69:1979–1985
Schmitt J, Hess H, Stunnenberg HG (1993) Affinity purification of histidine-tagged proteins. Mol Biol Rep 18:223–230
Seeman NC (2003) DNA in a material world. Nature 421:427–431
Takeda S, Tsukiji S, Nagamune T (2004) Site-specific conjugation of oligonucleotides to the C-terminus of recombinant protein by expressed protein ligation. Bioorg Med Chem Lett 14:2407–2410
Tominaga J, Kemori Y, Tanaka Y, Maruyama T, Kamiya N, Goto M (2007) An enzymatic method for site-specific labeling of recombinant proteins with oligonucleotides. Chem Commun 4:401–403
Tuerk C, Gold L (1990) Systematic evolution of ligands by exponential enrichment—RNA ligands to bacteriophage-T4 DNA-polymerase. Science 249:505–510
Tyagi S, Kramer FR (1996) Molecular beacons: probes that fluorescence upon hybridization. Nat Biotechnol 14:303–308
Yan H, Park SH, Finkelstein G, Reif JH, LaBean TH (2003) DNA-templated self-assembly of protein arrays and highly conductive nanowires. Science 301:1882–1884
Acknowledgements
The present work is supported by a Grant-in-Aid for the Global COE Program, “Science for Future Molecular Systems” from the Ministry of Education, Culture, Sports, Science and Technology of Japan.
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Shimada, J., Maruyama, T., Hosogi, T. et al. Conjugation of DNA with protein using His-tag chemistry and its application to the aptamer-based detection system. Biotechnol Lett 30, 2001–2006 (2008). https://doi.org/10.1007/s10529-008-9784-4
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DOI: https://doi.org/10.1007/s10529-008-9784-4