Large-scale production of soluble recombinant amyloid-β peptide 1-42 using cold-inducible expression system

被引:18
作者
Kim, Eun-Kyung [2 ]
Moon, Jeong Chan [2 ]
Lee, Jeong Mi [2 ]
Jeong, Min Seop [2 ]
Oh, Choongseob [1 ]
Ahn, Sung-Min [2 ]
Yoo, Yung Joon [1 ]
Jang, Ho Hee [2 ]
机构
[1] GIST, Sch Life Sci, Kwangju 500712, South Korea
[2] Gachon Univ, Lee Gil Ya Canc & Diabet Inst, Inchon 406799, South Korea
基金
新加坡国家研究基金会;
关键词
Aggregation-prone; Alzheimer's disease; Amyloid-beta peptide 1-42; Cell viability; Cold induction; Soluble expression; PROTEIN; AGGREGATION; PURIFICATION; CONVERSION; A-BETA-42;
D O I
10.1016/j.pep.2012.08.021
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Amyloid-beta peptide 1-42 (A beta(1-42)), the predominant form in senile plaques, plays important roles in the pathogenesis of Alzheimer's disease. Because A beta(1-42) has aggregation-prone nature, it has been difficult to produce in a soluble state in bacterial expression systems. In this study, we modified our expression system to increase the soluble fraction of A beta(1-42) in Escherichia coli (E. coli) cells. The expression level and solubility of recombinant A beta(1-42) induced at the low temperature (16 degrees C) is highly increased compared to that induced at 37 degrees C. To optimize expression temperature, the coding region of A beta(1-42) was constructed in a pCold vector, pCold-TF, which has a hexahistidine-tagged trigger factor (TF). Recombinant A beta(1-42) was expressed primarily as a soluble protein using pCold vector system and purified with a nickel-chelating resin. When the toxic effect of recombinant A beta(1-42) examined on human neuroblastoma SH-SY5Y cells, the purified A beta(1-42) induced cell toxicity on SH-SY5Y cells. In conclusion, the system developed in this study will provide a useful method for the production of aggregation prone-peptide such as A beta(1-42). (C) 2012 Elsevier Inc. All rights reserved.
引用
收藏
页码:53 / 57
页数:5
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