[Soluble expression of recombinant human apoliprotein A-I-Milano in Escherichia coli].

Abstract

Apolipoprotein A-I-Milano(AIM), a natural variant, not only inhibits the initiation and progression of atherosclerosis, but also makes the preexisting atherosclerotic lesions regress. AIM gene, at which N-terminal codens were optimized, was subcloned into the expression vector of pET22b. Recombiant plasmids were transformed into E. coli strain BL21 (DE3) and induced with IPTG. The expressed apoliprotein A-I-Milano was soluble in E. coli and was about 38% of total cell lysate. Purified by Butyl Sepharose 4F. F hydrophobic chromatography and Q Sepharose H.P. anion exchange chromatography, followed by ultrafiltration with Vivaspin 20 (30 000MW), AIM monomer was obtained in a purity of more than 95%. Activity assay of binding of AIM monomer to lipid indicates that association of AIM monomer with DMPC is slower than normal apoA-I but DMPC number associated by AIM monomer is more than by apoA-I. This results will be important for studying structure, function of AIM, specially clinical application.

• Authors: Ming Li, Hong-Liang Zhao, Chong Xue, Wei Zhang, Shi-Meng Zhang, Zhi-Min Liu
• Journal: Sheng wu gong cheng xue bao = Chinese journal of biotechnology (Sheng Wu Gong Cheng Xue Bao) Vol. 21 Issue 3 Pg. 354-9 (May 2005) ISSN: 1000-3061 [Print] China
• PMID: 16108355 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Apolipoprotein A-I
• Mutant Proteins
• Recombinant Proteins

Topics

• Apolipoprotein A-I (biosynthesis, genetics)
• Escherichia coli (genetics, metabolism)
• Humans
• Mutant Proteins (biosynthesis, genetics)
• Recombinant Proteins (biosynthesis, genetics)
• Solubility