- P-ISSN 2233-4203
- E-ISSN 2093-8950
- P-ISSN2233-4203
- E-ISSN2093-8950
- ESCI, SCOPUS, KCI
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Analysis of Phosphatidylinositol 3,4,5-Trisphosphates of PTEN Expression on Mammalian Cells
Mass Spectrometry Letters / Mass Spectrometry Letters, (P)2233-4203; (E)2093-8950
2013, v.4 no.3, pp.41-46
https://doi.org/10.5478/MSL.2013.4.3.41
(Konkuk University)
(Korea Basic Science Institute)
(Korea Basic Science Institute)
(Konkuk University)
(Konkuk University)
(Konkuk University)
(Konkuk University)
(Korea Basic Science Institute)
(Korea Basic Science Institute)
(Konkuk University)
(Konkuk University)
(Konkuk University)
(Konkuk University)
Jahan,
N., Park,
T., Kim,
Y. H., Lee,
D., Kim,
H., Noh,
K., &
Kim,
Y.
J.
(2013). Analysis of Phosphatidylinositol 3,4,5-Trisphosphates of PTEN Expression on Mammalian Cells. Mass Spectrometry Letters, 4(3), 41-46, https://doi.org/10.5478/MSL.2013.4.3.41
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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 cellularbehavior 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-stepextraction process using HCl for extraction of phosphatidylinositol 3,4,5-trisphosphates from two mammalian cell lines and furtheranalyzed the extracted phosphatidylinositol 3,4,5-trisphosphates using tandem mass spectrometry for the identification of them. Wefinally quantified the concentration of phosphatidylinositol 3,4,5-trisphosphates using internal standard calibration. From theseobservation, we found that HEK 293-T cells is a good model to examine the enzymatic behavior of PTEN in a cell, and theminimum 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 interms of the catalytic mechanism and further for the detailed identification of cellular substrates.
- keywords
- Phosphoinositides, Lipidomics, PTEN, Mass Spectrometry, Enzymatics
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- Submission Date
- 2013-07-22
- Revised Date
- 2013-09-24
- Accepted Date
- 2013-09-24
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