Mass Spectrometry Letters

Korean Society for Mass Spectrometry (한국질량분석학회)
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Analysis of Phosphatidylinositol 3,4,5-Trisphosphates of PTEN Expression on Mammalian Cells

  • Jahan, Nusrat (Department of Applied Biochemistry, Konkuk University) ;
  • Park, Taeseong (Division of Mass Spectrometry Research, Korea Basic Science Institute) ;
  • Kim, Young Hwan (Division of Mass Spectrometry Research, Korea Basic Science Institute) ;
  • Lee, Dongsun (Department of Applied Biochemistry, Konkuk University) ;
  • Kim, Hackyoung (Department of Applied Biochemistry, Konkuk University) ;
  • Noh, Kwangmo (Nanotechnology Research Center, Konkuk University) ;
  • Kim, Young Jun (Department of Applied Biochemistry, Konkuk University)
  • Received : 2013.07.22
  • Accepted : 2013.09.24
  • Published : 2013.09.30
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Abstract

The goal of this study is to find an experimental condition which enables us to perform enzymatic studies on the cellular behavior of PTEN (phosphatase and tensine homolog) through identification of molecular species of phosphatidylinositol 3,4,5-trisphosphates and their quantitative analysis in a mammalian cell line using mass spectrometry. We initially exployed a two-step extraction process using HCl for extraction of phosphatidylinositol 3,4,5-trisphosphates from two mammalian cell lines and further analyzed the extracted phosphatidylinositol 3,4,5-trisphosphates using tandem mass spectrometry for the identification of them. We finally quantified the concentration of phosphatidylinositol 3,4,5-trisphosphates using internal standard calibration. From these observation, we found that HEK 293-T cells is a good model to examine the enzymatic behavior of PTEN in a cell, and the minimum amount of phosphatidylinositol 3,4,5-trisphosphates is more than 50 pmol for quantification in a mass spectrometer. These results suggest that the well-optimized experimental conditions are required for the investigation of the cellular PTEN in terms of the catalytic mechanism and further for the detailed identification of cellular substrates.

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References

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