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.
Highlights
Type 2 diabetes mellitus (T2D), characterized by hyperglycemia, results from dysfunctional carbohydrate metabolism that is caused by a relative deficiency of insulin
Subsequent studies have led to conflicting claims about its physiological roles in promoting β cell proliferation [10, 11]. These findings suggested that angiopoietin-like protein 8 (ANGPTL8)/betatrophin was involved in triglyceride metabolism rather than glucose homeostasis [12]
Cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Gibco, USA) with 10% (v/v) fetal bovine serum (FBS), 2 mmol/L l-glutamine, 50 μmol/L 2-mercaptoethanol, 100 U/mL penicillin and 0.1 mg/mL streptomycin at a high (95%) relative humidity (RH) and 5% CO2 at 37 °C
Summary
Type 2 diabetes mellitus (T2D), characterized by hyperglycemia, results from dysfunctional carbohydrate metabolism that is caused by a relative deficiency of insulin. T2D has become a major health burden, with current prevalence estimated at 425 million and predicted to reach 629 million by 2045 worldwide [1]. The identification of novel molecules regulating glucose metabolism might provide new therapeutic targets for the treatment of T2D. Betatrophin, known as TD26 [2], refeeding induced fat and liver (RIFL) [3], lipoprotein lipase inhibition (lipasin) [4], and atypical angiopoietin-like protein 8 (ANGPTL8) [5], is primarily secreted by the human liver and white adipose tissue under the conditions of insulin resistance [6];. Bacteria‐Derived Recombinant Human ANGPTL8/Betatrophin Significantly Increases the Level
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