Production and Analysis of Biological Properties of Recombinant Human Apolipoprotein A-I

Abstract

Production of recombinant human apolipoprotein A-I (apoA-I) in E. coli cells is described and its biological properties are compared with those of natural protein. Recombinant apoA-I was isolated as a chimeric polypeptide and then processed to a mature form apoA-I (rapo-I). We studied the ability of the resulting protein to penetrate into hepatocyte nuclei and regulate the rate of DNA biosynthesis in complex with estriol. Penetration of rapoA-I conjugated with FITC into hepatocyte nuclei was demonstrated. rapoA-I-estriol and apoA-I-estriol complexes induced similar increase in DNA biosynthesis rate in isolated hepatocytes, which confi rms functional similarity of the obtained recombinant mature protein (rapoA-I) and native human apoA-I.

High‐efficient bacterial production of human ApoA‐I amyloidogenic variants

“…Of note, the yield of untagged WT‐ApoA‐I, obtained by using the method here described, is about 70% higher than the previously reported methods for the recombinant expression of native ApoA‐I, in both E. coli and Pichia pastoris. In addition, the yields of purified G26R‐ApoA‐I, L174S‐ApoA‐I, and L178H‐ApoA‐I variants, by using the same culture conditions and purification protocol, were determined to be in the same range as the WT protein (Table ), which indicates that the protocol can successfully be employed for amyloidogenic ApoA‐I variants in general.…”