Molecular Determinants for Protein Stabilization by Insertional Fusion to a Thermophilic Host Protein
| Autor: | Brennal Pierre, Edwin Aoraha, Jin Ryoun Kim, Asher Williams, Edward Chau, Jeffrey J. Gray, Tina Xiong, Jason W. Labonte, Kazi Yasin Helal, Vandan Shah |
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| Rok vydání: | 2015 |
| Předmět: |
Scaffold protein
Archaeal Proteins Entropy Recombinant Fusion Proteins Protein aggregation Biochemistry beta-Lactamases Protein–protein interaction Molecular Biology Protein Unfolding Calorimetry Differential Scanning biology Protein Stability Circular Dichroism Organic Chemistry Protein engineering biology.organism_classification Pyrococcus furiosus Kinetics Chaperone (protein) Chromatography Gel biology.protein Biophysics Molecular Medicine Target protein Protein stabilization |
| Zdroj: | ChemBioChem. 16:2392-2402 |
| ISSN: | 1439-4227 |
| DOI: | 10.1002/cbic.201500310 |
| Popis: | A universal method that improves protein stability and evolution has thus far eluded discovery. Recently, however, studies have shown that insertional fusion to a protein chaperone stabilized various target proteins with minimal negative effects. The improved stability was derived from insertion into a hyperthermophilic protein, Pyrococcus furiosus maltodextrin-binding protein (PfMBP), rather than from changes to the target protein sequence. In this report, by evaluating the thermodynamic and kinetic stability of various inserted β-lactamase (BLA) homologues, we were able to examine the molecular determinants of stability realized by insertional fusion to PfMBP. Results indicated that enhanced stability and suppressed aggregation of BLA stemmed from enthalpic and entropic mechanisms. This report also suggests that insertional fusion to a stable protein scaffold has the potential to be a useful method for improving protein stability, as well as functional protein evolution. |
| Databáze: | OpenAIRE |
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