Citation: Rongting Hu, Jue Wang, Jing Yang, Yu Ding, Guosong Chen. Micro Rings Self-assembled by Fluorescent Fusion-protein Based on Multiple Non-covalent Interactions[J]. Acta Polymerica Sinica, ;2019, 50(2): 135-146. doi: 10.11777/j.issn1000-3304.2018.18204 shu

Micro Rings Self-assembled by Fluorescent Fusion-protein Based on Multiple Non-covalent Interactions

Rongting Hu ^1,# ,
Jue Wang ^1,# ,
Jing Yang ¹ ,
Yu Ding ^2,, ,
Guosong Chen ^1,,

1.
Department of Macromolecular Science, Fudan University, Shanghai 200438
2.
Department of Physiology and Biophysics, School of Life Science, Fudan University, Shanghai 200438

Corresponding author: Yu Ding, yuding@fudan.edu.cn Guosong Chen, guosong@fudan.edu.cn
Received Date: 23 September 2018
Revised Date: 15 November 2018
Available Online: 4 January 2019

Proteins are attractive building blocks for construction of variant functional materials because of their chemical and structural diversities, and intrinsic functions. As the industry of biotechnology continues to expand, so does the expression of recombinant proteins with wide varieties. In this work, we adopted the recombinant protein technique to construct a new fusion protein, GFP-SA, as the building block of self-assemblies. The purification of GFP-SA was characterized by Superdex 75 size exclusive chromatography, SDS-PAGE, and MALDI-TOF. Then, GPC and native-PAGE were used to characterize the dimerization of GFP-SA based on the hydrogen bonds between neighboring SAs. Furthermore, ITC was employed to test the binding ability between GFP-SA and biotin, which revealed K_D = 0.24 μmol/L. In this study, we also designed and successfully synthesized the ligand RhYBio₂, which is composed of two biotin molecules and one rhodamine B molecule. The size of GFP-SA increased rapidly to 370 nm within one minute after mixing with RhYBio_2. We measured the particle size of GFP-SA/RhYBio₂ mixture every few minutes until the size stabilized at around 1300 nm 2 h later. However, size variation was barely observed for the controlled samples of SA/RhYBio2 (controlled protein) and GFP-SA/YBio (controlled ligand). We hypothesized that the two biotin molecules of RhYBio₂ could bind specifically with SA and align GFP-SA/RhYBio₂ into nanowires, which assembled further into micro rings. Their size was measured by dynamic light scattering (DLS) while the morphology was observed intuitively on a transmission electron microscope (TEM) and a confocal microscope (CM). The characteristic results from TEM and CM suggested an uneven size distribution of the micro rings prepared, which might be attributable to the flexibility of the fusion protein GFP-SA. These micro rings of GFP-SA/RhYBio₂ with fluorescence has great potential for biological applications.
- Fusion protein,
- GFP,
- Self-assembly,
- Micro rings
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