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In Vitro Assay to Measure Phosphatidylethanolamine Methyltransferase Activity
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Biologia
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JoVE Journal Biologia
In Vitro Assay to Measure Phosphatidylethanolamine Methyltransferase Activity

In Vitro Assay to Measure Phosphatidylethanolamine Methyltransferase Activity

DOI:
10.3791/53302-v

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09:33 min

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January 05, 2016

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1Department of Biological Sciences,St John’s University

Capitoli

  • 00:05Titolo
  • 00:58Cell Extract Preparation
  • 02:47Determination of the Protein Concentration of the Cell Extract Using Protein Estimation Kit
  • 03:46Enzymatic Assay
  • 05:14Lipid Extraction
  • 07:04Results: PEMT Activity is Specific and can be Measured from Whole Cell Extracts without the Need to Purify the Enzyme
  • 08:51Conclusion

Summary

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The present report describes an in vitro enzymatic assay to measure phosphatidylethanolamine methyltransferase activity using Leishmania cell extracts. This assay is based on the transfer of a radioactive methyl group from S-[Methyl-3H]adenosyl-L-methionine onto endogenous phosphatidylethanolamine.

Tags

Keywords: Phosphatidylethanolamine MethyltransferasePEMT ActivityLipid MetabolismPhosphatidylcholine BiosynthesisWhole Cell ExtractLeishmania CellsCell CultureCell LysisProtein QuantificationBCA Assay

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