Heterologous expression and purification of the soybean 7S globulin α′ subunit extension region: In vitro evidence of its involvement in cell cholesterol homeostasis

In a previous paper, the biological activity of a 216-amino acid recombinant truncated form of the soybean 7S globulin α′ subunit, known to control cholesterol and triglyceride homeostasis, was described. In this work, a shorter version of the polypeptide chain, spanning 142 amino acid residues from the N-terminus and thus exclusively including the so-called extension region, was cloned and overexpressed in Pichia pastoris. The yield of the recombinant polypeptide, which was termed α′E, was 8-fold greater than the previous truncated version. The α′E polypeptide was purified by simple conventional biochemical techniques to make it available for biological assays. Human hepatoma cell lines (Hep G2) were used to monitor the uptake and degradation of labeled low-density lipoproteins (LDL), according to an established procedure. The LDL uptake (+86%) and degradation (+94%) by cells tested at the highest α′E dose (2 μM) were similar to those found in cells incubated with 1 μM simvastatin, a potent inhibitor of cholesterol biosynthesis. Additionally, the cell response to α′E was found to be dose-dependent. The present findings strongly suggest that this recombinant polypeptide, or a fragment thereof, is the molecular determinant for cholesterol homeostasis and open new prospects for understanding the mechanism involved in this biological response, as a gateway to its utilization in lipid-lowering therapies.

► Soybean 7S globulin α′ subunit is known to control cholesterol and triglyceride homeostasis.
► The extension region of the α′ subunit (142 amino acid residues) was cloned and overexpressed in Pichia pastoris.
► Low density lipoprotein (LDL) uptake and degradation as affected by the recombinant polypeptide were measured in HepG2 cells.
► Both LDL uptake and degradation significantly increased in the cells treated with the recombinant soybean polypeptide.
► This finding opens the gateway to the development of a new drug in lipid-lowering therapies.