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Protein WISDOM: A Workbench for In silico De novo Design of BioMolecules
Published on: July 25, 2013
Bioinformatics-driven, rational engineering of protein thermostability.
Mary Kate Ditursi1, Seok-Joon Kwon, Philippa J Reeder
1Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute Troy, NY 12180-3590, USA.
Protein Engineering, Design & Selection : PEDS
|September 28, 2006
Summary
Protein engineering achieved enhanced thermophilicity by using bioinformatics to identify a key motif. Replacing a segment in subtilisin E increased its melting temperature and retained function at high temperatures.
Area of Science:
- Biochemistry
- Bioinformatics
- Protein Engineering
Background:
- Identifying sequence changes for desired protein functions is a key goal.
- Traditional methods include rational and random protein engineering.
- A bioinformatic approach offers an alternative for targeted protein modification.
Purpose of the Study:
- To identify specific sequence changes influencing protein functional properties.
- To pinpoint a motif contributing to thermophilicity in the serine protease subtilase superfamily.
- To engineer a thermophilic variant of subtilisin E.
Main Methods:
- Employed Bayesian sequence-based algorithms (PROBE and Classifier).
- Identified a strand-turn-strand motif associated with thermophilicity.
- Replaced a 16 amino acid sequence in mesophilic subtilisin E with a modeled thermophilic sequence.
Main Results:
- The engineered subtilisin E exhibited a 13°C increase in melting temperature.
- The mutant enzyme retained significant function at 90°C, unlike the wild-type.
- The modified subtilisin E retained approximately one-third of its activity at 45°C compared to the wild-type.
Conclusions:
- Bioinformatic analysis can effectively identify sequence motifs that enhance protein thermophilicity.
- Targeted sequence modification based on computational predictions can significantly improve protein stability and function at elevated temperatures.
- This approach provides a powerful tool for protein engineering, enabling the design of enzymes with tailored properties for specific applications.
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