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Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures
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| Zeitschriftentitel: | Chemistry – A European Journal |
|---|---|
| Personen und Körperschaften: | , , , |
| In: | Chemistry – A European Journal, 25, 2019, 14, S. 3483-3488 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Wiley
|
| Schlagwörter: |
| author_facet |
Burgahn, Teresa Garrecht, Ruben Rabe, Kersten S. Niemeyer, Christof M. Burgahn, Teresa Garrecht, Ruben Rabe, Kersten S. Niemeyer, Christof M. |
|---|---|
| author |
Burgahn, Teresa Garrecht, Ruben Rabe, Kersten S. Niemeyer, Christof M. |
| spellingShingle |
Burgahn, Teresa Garrecht, Ruben Rabe, Kersten S. Niemeyer, Christof M. Chemistry – A European Journal Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures General Chemistry Catalysis Organic Chemistry |
| author_sort |
burgahn, teresa |
| spelling |
Burgahn, Teresa Garrecht, Ruben Rabe, Kersten S. Niemeyer, Christof M. 0947-6539 1521-3765 Wiley General Chemistry Catalysis Organic Chemistry http://dx.doi.org/10.1002/chem.201805506 <jats:title>Abstract</jats:title><jats:p>We present a facile method for the combined synthesis and purification of protein‐decorated DNA origami nanostructures (DONs). DONs bearing reductively cleavable biotin groups in addition to ligands for ligation of recombinant proteins are bound to magnetic beads. Protein immobilization is conducted with a large protein excess to achieve high ligation yields. Subsequent to cleavage from the solid support, pure sample solutions are obtained which are suitable for direct AFM analysis of occupation patterns. We demonstrate the method's utility using three different orthogonal ligation methods, the “halo‐based oligonucleotide binder” (HOB), a variant of Halo‐tag, the “SpyTag/SpyCatcher” (ST/SC) system, and the enzymatic “ybbR tag” coupling. We find surprisingly low efficiency for ST/SC ligation, presumably due to electrostatic repulsion and steric hindrance, whereas the ybbR method, despite its ternary nature, shows good ligation yields. Our method is particularly useful for the development of novel ligation methods and the synthesis of mechanically fragile DONs that present protein patterns for surface‐based cell assays.</jats:p> Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures Chemistry – A European Journal |
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| title |
Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures |
| title_unstemmed |
Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures |
| title_full |
Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures |
| title_fullStr |
Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures |
| title_full_unstemmed |
Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures |
| title_short |
Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures |
| title_sort |
solid‐phase synthesis and purification of protein–dna origami nanostructures |
| topic |
General Chemistry Catalysis Organic Chemistry |
| url |
http://dx.doi.org/10.1002/chem.201805506 |
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2019 |
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3483-3488 |
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<jats:title>Abstract</jats:title><jats:p>We present a facile method for the combined synthesis and purification of protein‐decorated DNA origami nanostructures (DONs). DONs bearing reductively cleavable biotin groups in addition to ligands for ligation of recombinant proteins are bound to magnetic beads. Protein immobilization is conducted with a large protein excess to achieve high ligation yields. Subsequent to cleavage from the solid support, pure sample solutions are obtained which are suitable for direct AFM analysis of occupation patterns. We demonstrate the method's utility using three different orthogonal ligation methods, the “halo‐based oligonucleotide binder” (HOB), a variant of Halo‐tag, the “SpyTag/SpyCatcher” (ST/SC) system, and the enzymatic “ybbR tag” coupling. We find surprisingly low efficiency for ST/SC ligation, presumably due to electrostatic repulsion and steric hindrance, whereas the ybbR method, despite its ternary nature, shows good ligation yields. Our method is particularly useful for the development of novel ligation methods and the synthesis of mechanically fragile DONs that present protein patterns for surface‐based cell assays.</jats:p> |
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| author | Burgahn, Teresa, Garrecht, Ruben, Rabe, Kersten S., Niemeyer, Christof M. |
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| description | <jats:title>Abstract</jats:title><jats:p>We present a facile method for the combined synthesis and purification of protein‐decorated DNA origami nanostructures (DONs). DONs bearing reductively cleavable biotin groups in addition to ligands for ligation of recombinant proteins are bound to magnetic beads. Protein immobilization is conducted with a large protein excess to achieve high ligation yields. Subsequent to cleavage from the solid support, pure sample solutions are obtained which are suitable for direct AFM analysis of occupation patterns. We demonstrate the method's utility using three different orthogonal ligation methods, the “halo‐based oligonucleotide binder” (HOB), a variant of Halo‐tag, the “SpyTag/SpyCatcher” (ST/SC) system, and the enzymatic “ybbR tag” coupling. We find surprisingly low efficiency for ST/SC ligation, presumably due to electrostatic repulsion and steric hindrance, whereas the ybbR method, despite its ternary nature, shows good ligation yields. Our method is particularly useful for the development of novel ligation methods and the synthesis of mechanically fragile DONs that present protein patterns for surface‐based cell assays.</jats:p> |
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| spelling | Burgahn, Teresa Garrecht, Ruben Rabe, Kersten S. Niemeyer, Christof M. 0947-6539 1521-3765 Wiley General Chemistry Catalysis Organic Chemistry http://dx.doi.org/10.1002/chem.201805506 <jats:title>Abstract</jats:title><jats:p>We present a facile method for the combined synthesis and purification of protein‐decorated DNA origami nanostructures (DONs). DONs bearing reductively cleavable biotin groups in addition to ligands for ligation of recombinant proteins are bound to magnetic beads. Protein immobilization is conducted with a large protein excess to achieve high ligation yields. Subsequent to cleavage from the solid support, pure sample solutions are obtained which are suitable for direct AFM analysis of occupation patterns. We demonstrate the method's utility using three different orthogonal ligation methods, the “halo‐based oligonucleotide binder” (HOB), a variant of Halo‐tag, the “SpyTag/SpyCatcher” (ST/SC) system, and the enzymatic “ybbR tag” coupling. We find surprisingly low efficiency for ST/SC ligation, presumably due to electrostatic repulsion and steric hindrance, whereas the ybbR method, despite its ternary nature, shows good ligation yields. Our method is particularly useful for the development of novel ligation methods and the synthesis of mechanically fragile DONs that present protein patterns for surface‐based cell assays.</jats:p> Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures Chemistry – A European Journal |
| spellingShingle | Burgahn, Teresa, Garrecht, Ruben, Rabe, Kersten S., Niemeyer, Christof M., Chemistry – A European Journal, Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures, General Chemistry, Catalysis, Organic Chemistry |
| title | Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures |
| title_full | Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures |
| title_fullStr | Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures |
| title_full_unstemmed | Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures |
| title_short | Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures |
| title_sort | solid‐phase synthesis and purification of protein–dna origami nanostructures |
| title_unstemmed | Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures |
| topic | General Chemistry, Catalysis, Organic Chemistry |
| url | http://dx.doi.org/10.1002/chem.201805506 |