A New Focus on Prone Peptide Purification in the Form of Inclusion Bodies and the Advantage of Using Deubiquitylating Enzyme in Purifying Those Peptides
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
Abstract 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
Merlin Jayalal .L .P [ Department of Biochemistry, Bharathidasan college of Arts and Science, Ellispettai, Erode-638116, Tamilnadu, India ]
Corresponding author
조선대학교 기초과학연구원 [The Natural Science Research Institute of Chosun]
설립연도
2008
분야
자연과학>자연과학일반
소개
본 연구원은 기초과학을 진흥하기 위한 연구·교육 및 그 보급을 목적으로 한다. 이 목적을 달성하기 위하여 다음 각 호의 사업을 수행한다.
1. 기초과학 제 분야에 관한 조사와 연구
2. 기초과학에 관한 학술행사(학술대회, 학술세미나, 심포지엄, 초청강연회 등) 개최
3. 학문후속세대 및 일반인을 위한 기초과학 교육
4. 기관지『조선자연과학논문지』 발간
5. 『자연과학연구총서』, 『자연과학번역총서』 등 단행본 발간
6. 기타 본 연구원의 목적과 관련된 사업
간행물
간행물명
통합자연과학논문집(구 조선자연과학논문집) [Journal of Integrative Natural Science]