CHEMISTRY OF BIOCONJUGATES
Semisynthetic DNA-protein conjugates are synthesized by either covalent coupling chemistry, or by means of reversible biomolecular recognition systems. For instance, efficient syntheses of covalent conjugates of the biotin-binding protein streptavidin, enzymes and antibodies with single-stranded DNA oligomers are being developed. Experimental methods involved include nucleic acid and pepide synthesis, development of reactive linker compounds, the expression of recombinant proteins and other techniques of modern bioorganic chemistry and molecular biology.
A second approach to semisynthetic DNA-protein conjugates takes advantage of non-covalent self-assembly based on biomolecular recognition systems. In particular, the hybridization of complementary nucleic acids and the high-affinity binding of biotin and streptavidin are utilized to generate supramolecular bioconjugates. Examples include the controlled formation of oligomeric conjugates from end-biotinylated DNA fragments and streptavidin, which can be converted to well-defined supramolecular nanocircles. These type of DNA-streptavidin conjugates are applicable as reagents in Immunological Diagnostics and as functional building blocks for the generation of DNA Nanostructures.
Analytical methods for the characterization of the semisynthetic DNA-protein conjugates include chromatographic and electrophoretic techniques, SPR affinity measurements, mass spectrometry, electron and scanning-probe microscopy.
References:
- Lovrinovic, M., Spengler, M., Deutsch, C., Niemeyer, C. M. (2005) Synthesis of covalent DNA-protein conjugates by expressed protein ligation. Mol. Biosys. 1, 64-69.
- Fruk, L., Niemeyer, C. M. (2005) Covalent Hemin-DNA Adducts for Generating a Novel Class of Artifical Heme Enzymes. Angew. Chem. Int. Ed. 44, 2603 -2606; Angew. Chem. 2005, 117, 2659-2662.
- Kukolka, F., Niemeyer, C. M. (2004) Synthesis of fluorescent oligonucleotide-EYFP conjugate: Towards supramolecular construction of semisynthetic biomolecular antennae. Org. Biomol. Chem. 2, 2203 - 2206
- Niemeyer, C. M. (Ed.) Bioconjugation Protocols: Strategies and Methods, in Methods in Molecular Biology, Humana Press, Totowa, New Jersey 2004, ISBN 1-58829-098-0.
- Lovrinovic, M., Seidel, R., Wacker, R., Schroeder, H., Seitz, O., Engelhard, M., Goody, R., Niemeyer, C. M. (2003) Synthesis of protein-nucleic acid conjugates by expressed protein ligation. Chem. Commun. 822-823.
- Richter, J., Adler, M., Niemeyer, C. M. (2003) Monte-Carlo-Simulation of the Assembly of Bis-Biotinylated DNA and Streptavidin. ChemPhysChem 4, 79-83.
- Niemeyer, C. M., Adler, M., Gao, S., Chi, L. F. (2002) Supramolecular DNA-Streptavidin Nanocircles with a of Covalently Attached Oligonucleotide Moiety; J. Biomol. Struct. Dyn., 223-230.
- Niemeyer, C. M., Koehler, J., Wuerdemann, C. (2002) DNA-Directed Immobilization of an Artificial Bienzyme Consisting of NAD(P)H:FMN Oxidoreductase and Luciferase; ChemBioChem, 3, 242-245.
- Pignataro, B., Chi, L. F., Gao, S., Anczykowsky, B., Niemeyer, C. M., Adler, M., Fuchs, H. (2002) Dynamic Scanning Force Microscopy Study of Self-Assembled DNA-Protein Oligomers. Appl. Phys. A, 74, 447-452.
- Niemeyer, C. M., Adler, M., Gao, S., Chi, L. F. (2001) Nanostructured DNA-Protein Aggregates Consisting of Covalent Oligonucleotide-Streptavidin Conjugates. Bioconjug. Chem., 12, 364-371.
- Niemeyer, C. M., Adler, M., Gao, S., Chi, L. F. (2000) Supramolecular Nanocircles Consisting of Streptavidin and DNA. Angew. Chem. 112, 3183-3187, Angew. Chem. Int. Ed. 39, 3055-3059.
- Niemeyer, C. M., Ceyhan, B., Blohm, D. (1999) Functionalization of Covalent DNA-Streptavidin Conjugates by Means of Biotinylated Modulator Components. Bioconjugate Chem. 10, 708-719.
- Niemeyer, C. M., Bürger, W., Hoedemakers, R. M. J. (1998) Hybridization Characteristics of Biomolecular Adaptors, Covalent DNA-streptavidin Conjugates. Bioconjugate Chem. 9, 168-175 .
- Niemeyer, C. M., Sano, T., Smith, C. L., Cantor, C. R. (1994) Oligonucleotide-directed self-assembly of proteins: Semisynthetic DNA-streptavidin hybrid molecules as connectors for the generation of macroscopic arrays and the construction of supramolecular bioconjugates. Nucleic Acids Res. 22, 5530-5539 .
