Efficient Formation of Site-Specific Protein-DNA Hybrids Using Copper-Free Click Chemistry.
Protein-DNA hybrids have become increasingly popular molecular building blocks in bionanotechnology and single-molecule studies to synergistically combine the programmability of DNA with the chemical diversity of proteins. The growing demand for protein-DNA hybrids requires powerful strategies for their conjugation. Here, we present an efficient two-step method for protein-DNA assembly based on copper-free click chemistry. The method allows site-specificity and high coupling efficiency, while maintaining the conservation of protein activity. We compare our method to a commonly used protocol of direct linkage of maleimide-modified oligos. We demonstrate the significantly higher yield with a protein-DNA conjugate, which is analyzed using single-molecule force spectroscopy.
B CUBE - Center for Molecular Bioengineering, Technische Universität Dresden , Arnoldstraße 18, 01307 Dresden, Germany.
Ann
A
Mukhortava
B CUBE - Center for Molecular Bioengineering, Technische Universität Dresden , Arnoldstraße 18, 01307 Dresden, Germany.
Michael
M
Schlierf
D001386:Azides / Q000737:chemistry; D057930:Click Chemistry; D004247:DNA / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D008958:Models, Molecular; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*
D001386:Azides; D011506:Proteins; D004247:DNA