Insights into Vitamin D metabolism using cyp24 over-expression and knockout systems in conjunction with liquid chromatography/mass spectrometry (LC/MS)

The Journal of Steroid Biochemistry and Molecular Biology
Sonoko MasudaGlenville Jones

Abstract

The development of novel gene expression systems for cytochrome P450s (CYPs) together with a revolution in analytical mass spectrometry with the emergence of liquid chromatography/mass spectrometry (LC/MS) has opened the door to answering some long-standing questions in Vitamin D metabolism. Our studies focused on: (1) elucidating the role of CYP24 in 25-OH-D3 and 1alpha,25-(OH)2D3 metabolism; (2) exploring how DBP influences this process; (3) measuring 25-OH-D3 metabolism in CYP24-knockout (CYP24-XO) cells and; (4) comparing 1alpha-OH-D2 metabolism in the CYP24-XO mouse in vivo and in vitro. Methodology employed CYP24 over-expression and knockout systems in conjunction with state-of-the-art analytical LC/MS, diode array, and radioisotopic detection methods. We found that CYP24 metabolizes 25-OH-D3 and 1alpha,25-(OH)2D3 at similar rates in vitro, but that for 25-OH-D3 but not 1alpha,25-(OH)2D3, this rate is strongly influenced by the concentration of DBP. Unlike their wild type littermates, the administration of 25-OH-D3 to CYP24-XO mice results in no measurable 24,25-(OH)2D3 production. When neonatal murine keratinocytes are prepared from wild type and CYP24-XO mice there was no measurable production of 24,25-(OH)2D3 or 1alpha...Continue Reading

References

Jan 10, 2001·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·T TaniguchiT Setoguchi
Jan 5, 2002·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·Glenville Jones, Harriet S Tenenhouse
Jan 10, 2003·Journal of Cellular Biochemistry·R L HorstS Reddy

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Citations

Aug 1, 2015·Nutrients·Noor H A SuainiLeonard C Harrison
Aug 19, 2016·Journal of the American Society of Nephrology : JASN·Loan Nguyen-YamamotoDavid Goltzman
Jun 5, 2014·The FEBS Journal·Elaine W TieuRobert C Tuckey
Oct 26, 2016·Oncology Reports·Makoto Osanai, Gang-Hong Lee
Jul 12, 2019·Frontiers in Immunology·Juan Navarro-BarriusoEva M Martínez-Cáceres
Jan 27, 2005·Journal of Mass Spectrometry : JMS

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