Chemoenzymatic synthesis of glutamate mutase S component
| Autor: | Heng-Ju Lin, 林恆如 |
|---|---|
| Rok vydání: | 2005 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 93 Glutamate mutase S component (MutS) is the smallest known protein subunit that carries cobalamin-binding domain with molecular masses of 14,748 Da. The chemo-enzymatic synthesis of this small monomeric protein by the native chemical ligation method is the long-term goal of in this study. Two unprotected peptide segment, 1 and 2, were synthesized separately. The segment 1 (residue 1-25) was readily prepared in good yield by solid-phase peptide synthesis. A thio-ester group was added to the carboxyl end of the segment 1. The segment 2 (residue 26-131) was produced by the over-expression of an engineered and truncated mutS gene. After treatment with TEV protease to remove the hexa-His purification tag, the segment 2 peptide carries a cysteine group at its N-terminal end. The ligation of above two fragments a rapid intramolecular S→N acyl shift through will be subsequently carried out in our group. This result indicates that incorporation of an artificial amino acid residue into the conserved cobalamin-binding motif is feasible by using this synthetic strategy. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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