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Synthesis of proteins by subtiligase
A C Braisted1, J K Judice, J A Wells
1Department of Protein Engineering, Genentech, Inc., South San Francisco, California 94080, USA.
Methods in Enzymology
|January 1, 1997
Summary
Protein engineering created subtiligase, an efficient peptide ligase for protein synthesis. This novel enzyme shows promise for synthesizing proteins via segment condensation, including RNase A.
Area of Science:
- Biochemistry
- Protein Engineering
- Enzymology
Background:
- Subtilisin, a serine protease, has been a target for protein engineering.
- Developing efficient methods for protein synthesis, particularly via segment condensation, is crucial.
Purpose of the Study:
- To engineer an efficient peptide ligase from subtilisin.
- To explore the applications of the novel enzyme, subtiligase, in protein synthesis.
Main Methods:
- Utilized protein engineering strategies to redesign the active site of subtilisin.
- Applied the engineered enzyme, subtiligase, for protein synthesis and semisynthesis.
Main Results:
- Successfully generated subtiligase, an efficient peptide ligase.
- Demonstrated the utility of subtiligase in the total synthesis of RNase A and semisynthesis of other proteins.
Conclusions:
- Subtiligase is a promising novel enzyme for protein synthesis.
- This engineered peptide ligase offers a valuable addition to protein segment condensation strategies.