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Click-Chemistry Based Fluorometric Assay for Apolipoprotein N-acyltransferase from Enzyme Characterization to High-Throughput Screening
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生物化学
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JoVE 杂志 生物化学
Click-Chemistry Based Fluorometric Assay for Apolipoprotein N-acyltransferase from Enzyme Characterization to High-Throughput Screening

Click-Chemistry Based Fluorometric Assay for Apolipoprotein N-acyltransferase from Enzyme Characterization to High-Throughput Screening

DOI:
10.3791/61146-v

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07:37 min

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May 13, 2020

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, ,
1Institut Pasteur, Unité Biologie et génétique de la paroi bactérienne, Paris, France ; CNRS, UMR 2001 « Microbiologie intégrative et Moléculaire », Paris, France ; INSERM, Équipe Avenir, Paris, France

Chapters

  • 00:05Introduction
  • 00:55Tube Assay
  • 05:23Results: Fluorometric Enzyme Assay for Apolipoprotein N-Acyltransferase
  • 07:09Conclusion

Summary

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Presented here is a sensitive fluorescence assay to monitor apolipoprotein N-acyltransferase activity using diacylglyceryl peptide and alkyne-phospholipids as substrates with click-chemistry.

Tags

Click-chemistryFluorometric AssayApolipoprotein N-acyltransferaseEnzyme CharacterizationHigh-throughput ScreeningIntegral Membrane EnzymeLipidDetergentSonicationAlkyne POPEFSL-1-biotinLNT Reaction BufferStreptavidinAzido-FAMTCEPTBTACopper SulfatePBST1Nonhomogeneous SuspensionPotential Antibiotics

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