Prokaryotic expression of human vitamin D receptor (hVDR) and its binding activities to Cd and Pb

• Published in: Huanjing kexue Vol. 31; no. 10; p. 2469
• Main Authors: Jiao, Jian, Zhang, Zhao-Bin, Hu, Jian-Ying, Wang, Jin-Shui
• Format: Journal Article
• Language: Chinese
• Published: China 01.10.2010
• Subjects: Alkaline Phosphatase - metabolism; Cadmium - metabolism; Cadmium - toxicity; Cloning, Molecular; Environmental Pollutants - metabolism; Environmental Pollutants - toxicity; Genetic Vectors - genetics; Humans; Lead - metabolism; Lead - toxicity; Nuclear Receptor Coactivator 2 - metabolism; Osteoporosis - chemically induced; Receptors, Calcitriol - biosynthesis; Receptors, Calcitriol - genetics; Receptors, Calcitriol - metabolism; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - genetics
• ISSN: 0250-3301

Human vitamin D receptor (hVDR) is a potential receptor channel for heavy metals to affect bone metabolism, while to date there is no report about the binding activity between heavy metals and hVDR. This study established a prokaryotic expression of hVDR system, by cloned hVDR-LBD into pGEX-4T-1 vector. Then according to the principle that the nuclear receptor binding with its ligand can be combined with nuclear receptor coactivator 2-bacterial alkaline phosphatase fusion protein (TIF2-BAP), we established a method of p-nitrophenylphosphate-alkaline phosphatase to analyse the effects of chemical on the binding activity between hVDR and TIF2-BAP. Using this method, we studied the binding activities between hVDR and TIF2-BAP after exposure of cadmium and lead. The results showed that the binding activities significantly increased to 3.95 and 4.39 times that of the control after exposure of 100 micromol/L and 1 000 micromol/L cadmium chloride (CdCl2), and the binding activities significantly increased to 2.29 an