Tuning Selective Transport of Biomolecules through Site-Mutated Nucleoporin-like Protein (NLP) Hydrogels
| Autor: | Yun Jung Yang, Melody Morris, Shuaili Li, Danielle J. Mai, Bradley D. Olsen |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Nucleocytoplasmic Transport Proteins
Saccharomyces cerevisiae Proteins Polymers and Plastics Active Transport Cell Nucleus Bioengineering Sequence (biology) Saccharomyces cerevisiae 02 engineering and technology 010402 general chemistry computer.software_genre 01 natural sciences Biomaterials Materials Chemistry medicine Nuclear pore Cell Nucleus chemistry.chemical_classification business.industry Biomolecule Nuclear Proteins Hydrogels 021001 nanoscience & nanotechnology 0104 chemical sciences Nuclear Pore Complex Proteins Cell nucleus medicine.anatomical_structure chemistry Self-healing hydrogels Nucleoporin Artificial intelligence Nuclear transport 0210 nano-technology business computer Natural language processing Function (biology) |
| Zdroj: | Biomacromolecules. 22:289-298 |
| ISSN: | 1526-4602 1525-7797 |
| DOI: | 10.1021/acs.biomac.0c01083 |
| Popis: | Natural selective filtering systems (e.g., the extracellular matrix, nuclear pores, and mucus) separate molecules selectively and efficiently, and the detailed understanding of transport mechanisms exploited in these systems provides important bioinspired design principles for selective filters. In particular, nucleoporins consist of consensus repeat sequences that are readily utilized for engineering repeat proteins. Here, the consensus repeat sequence of Nsp1, a yeast nucleoporin, is polymerized to form a nucleoporin-like protein (NLP) and mutated to understand the effect of sequence on selective transport. The hydrophilic spacers of the NLPs were redesigned considering net charge, charge distribution, and polarity. Mutations were made near to and far from the FSFG interacting domain to explore the role of highly conserved residues as a function of spatial proximity. A nuclear transport receptor-cargo complex, nuclear transport factor 2-green fluorescent protein (NTF2-GFP), was used as a model for changes in transport. For mutations of the charged spacer, some mutations of highly conserved charged residues were possible without knocking out selective transport of the NTF2, but the formation of regions of clustered negative charge has an unfavorable effect on nuclear transporter permeation. Thus, positive net charge and alternating positive and negative charge within the hydrophilic spacer are advantageous for recognition and selective transport. In the polarity panel, mutations that increased the interaction between NTF2-GFP and the gel led to decreased permeation of the NTF2-GFP due to blocking of the interface and inability of the NTF2-GFP to transport into the gel. Therefore, these results provide a strategy for tuning selective permeability of biomolecules using the artificially designed consensus repeat-based hydrogels. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|