| Chapter title |
Enzyme Encapsulation in an Engineered Lumazine Synthase Protein Cage
|
|---|---|
| Chapter number | 4 |
| Book title |
Protein Scaffolds
|
| Published in |
Methods in molecular biology, January 2018
|
| DOI | 10.1007/978-1-4939-7893-9_4 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7892-2, 978-1-4939-7893-9
|
| Authors |
Yusuke Azuma, Donald Hilvert, Azuma, Yusuke, Hilvert, Donald |
| Abstract |
The packaging of active enzymes in protein cages is a powerful strategy to control catalytic activity. Using a positively supercharged variant of green fluorescent protein, GFP(+36), as a genetically programmable tag, enzymes can be rapidly and quantitatively loaded into an engineered variant of the Aquifex aeolicus cage-forming protein lumazine synthase (AaLS-13) that possesses a negatively charged lumen. The cargo is spontaneously localized within AaLS-13 cages by simply mixing the components in aqueous solution. This chapter describes a detailed protocol for the preparation of AaLS-13 cages and GFP(+36)-enzyme fusions, as well as characterization of the inclusion complexes. Suitable conditions for encapsulation and enzyme kinetic assays are also discussed. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 25 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 5 | 20% |
| Student > Bachelor | 4 | 16% |
| Student > Master | 2 | 8% |
| Student > Ph. D. Student | 1 | 4% |
| Other | 1 | 4% |
| Other | 1 | 4% |
| Unknown | 11 | 44% |
| Readers by discipline | Count | As % |
|---|---|---|
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| Chemical Engineering | 1 | 4% |
| Pharmacology, Toxicology and Pharmaceutical Science | 1 | 4% |
| Psychology | 1 | 4% |
| Medicine and Dentistry | 1 | 4% |
| Other | 2 | 8% |
| Unknown | 11 | 44% |