원문정보
초록
영어
Synthesis of long or aggregation-prone peptide has been problematic chemically. Its biological production has an advantage in that point, but it often forms inclusion body which creates difficulties in recovery of targets. As a deubiquitylating enzyme (Usp2-cc) was shown in this study to maintain its activity even in the presence of up to 4 M urea, target peptide was purified by a single step of chromatography after overexpression as inclusion body, solubilization in urea and cleavage by the enzyme from the fusion protein consisting of GroES (used for high expression and easy to handle), ubiquitin (as a cleavage site) and target peptide. This system is a convenient tool for production of peptides that are difficult to be chemically synthesized and biologically purified.
목차
1. Introduction
2. Experimental Section
2.1. Materials
2.2. Construction of Vector for the Fusion Protein
2.3. Expression of GroES Fusion Proteins, Purification of Peptides and Usp2-cc
2.4. Cell Culture and Biological Effects
2.5. Secondary Structure Analysis of Aβ1–42
2.6. Fibrillogenesis of Ab1–42
3. Results and Discussion
3.1. Construction of Vector for a High Level Expression
3.2. Cleavage of Fusion Protein in Moderate Concentrations of Urea
3.3. Purification and Characterization of Peptides
4. Conclusion
References