Bacteria-Derived Recombinant Human ANGPTL8/Betatrophin Significantly Increases the Level of Triglyceride

Protein J. 2019 Aug;38(4):472-478. doi: 10.1007/s10930-019-09825-8.

Abstract

ANGPTL8/Betatrophin has been implicated in the regulation of both glucose and triglyceride metabolism. However, its role in regulating glucose metabolism by promoting β cell proliferation remains controversial, and its physiological functions and molecular targets are largely unknown. Hence, it is of great importance to make recombinant protein and test its effects on β cell mass directly. In this study, the mature form gene of human ANGPTL8/betatrophin was obtained through chemical synthesis on to the vector pUCE, and the fusion protein was expressed in the Transetta (DE3)/pEASY-E2-betatrophin strain. The inclusion bodies were solubilized in urea and purified by Ni-NTA affinity chromatography. The yield of purified ANGPTL8/betatrophin was approximately 20 mg per liter of culture medium. In vitro studies revealed that the recombinant ANGPTL8/betatrophin had no proliferation effect on MIN6 cells but promoted TG levels in HepG2 cells. This method to generate bioactive ANGPTL8/betatrophin is a simple, practical and user-friendly protocol.

Keywords: ANGPTL8/betatrophin; Beta cell proliferation; Recombinant expression; Triglyceride level.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Angiopoietin-Like Protein 8
  • Angiopoietin-like Proteins / isolation & purification*
  • Angiopoietin-like Proteins / pharmacology*
  • Cell Proliferation / drug effects*
  • Cloning, Molecular
  • Escherichia coli / genetics
  • Hep G2 Cells
  • Humans
  • Peptide Hormones / isolation & purification*
  • Peptide Hormones / pharmacology*
  • Recombinant Proteins / pharmacology*
  • Triglycerides / metabolism*

Substances

  • ANGPTL8 protein, human
  • Angiopoietin-Like Protein 8
  • Angiopoietin-like Proteins
  • Peptide Hormones
  • Recombinant Proteins
  • Triglycerides