| Autor: |
Mickoleit F; Dept. Microbiology, University of Bayreuth, D-95447 Bayreuth, Germany., Beierl JJ; Dept. Microbiology, University of Bayreuth, D-95447 Bayreuth, Germany., Markert S; Dept. Microbiology, University of Bayreuth, D-95447 Bayreuth, Germany., Klein MA; Dept. Microbiology, University of Bayreuth, D-95447 Bayreuth, Germany., Stäbler SY; Dept. Microbiology, University of Bayreuth, D-95447 Bayreuth, Germany., Maier DS; Dept. Microbiology, University of Bayreuth, D-95447 Bayreuth, Germany., Schüler D; Dept. Microbiology, University of Bayreuth, D-95447 Bayreuth, Germany. |
| Abstrakt: |
Bacterial magnetosomes ("MAGs") represent a promising class of magnetic iron oxide nanoparticles with exceptional material characteristics and high application potential in the biomedical and biotechnological field. For the surface functionalization of MAGs with different protein cargos, their enveloping membrane can be addressed by genetic means. However, the expression of foreign polypeptides as translational fusion to magnetosome membrane proteins is still laborious and lacks versatility as the generated particles are monospecific and thus restricted to predetermined functions. Utilizing the SpyTag-SpyCatcher (ST-SC) bioconjugate system, we here establish a flexible platform for the targeted nanoassembly of multifunctional MAGs that combines the rapidity of chemical coupling (e.g., by cross-linking reactions) and the unmatched selectivity and controllability of in vivo functionalization. MAGs genetically engineered to display either SC- or ST-connectors are shown to efficiently bind a variety of complementary tagged (protein) cargo. Specifically, we cover a broad spectrum of representative functional moieties and foreign cargo (such as enzymes, antibodies, fluorophores, and silica beads) with relevance in biotechnology and biomedicine and demonstrate the interchangeability of the MAGs-adapted ST-SC system. For the controlled generation of artificial shells surrounding the particles, SC-MAGs are effectively coated by protein-corona proteins. The potential of the here-provided toolkit is even more enhanced by using SC-MAGs as an affinity tool for selective protein pulldown in vitro and in vivo . Overall, this innovative technology turns bacterial MAGs into a flexible magnetic nanoscaffold for the targeted plug-and-play display of virtually unlimited additional functionalities, thereby generating a multitude of magnetic hybrid materials that can be used in many applications. |
