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Profiling of Methyltransferases and Other S-adenosyl-L-homocysteine-binding Proteins by Capture Compound Mass Spectrometry (CCMS)
Journal JoVE
Biologie
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Journal JoVE Biologie
Profiling of Methyltransferases and Other S-adenosyl-L-homocysteine-binding Proteins by Capture Compound Mass Spectrometry (CCMS)

Profiling of Methyltransferases and Other S-adenosyl-L-homocysteine-binding Proteins by Capture Compound Mass Spectrometry (CCMS)

DOI:
10.3791/2264-v

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17:12 min

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December 20, 2010

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, , , , , ,
1Department of Biochemistry / Analytics,caprotec bioanalytics GmbH, 2Institute of Organic Chemistry,RWTH Aachen University

Chapitres

  • 00:05Titre
  • 02:23Performing the Capture Assay and Controls
  • 08:26Tryptic Digest of Captured Proteins and Preparation of Peptides for LC-MS/MS
  • 09:50Peptide and Protein Identification via Automated Sequence Database Search
  • 12:33Representative Results of CCMS Experiments
  • 16:00Conclusion

Summary

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Traduction automatique

Capture Compounds are trifunctional small molecules to reduce the complexity of the proteome by functional reversible small molecule-protein interaction followed by photo-crosslinking and purification. Here we use a Capture Compound with S-adenosyl-L-homocysteine-binding as selectivity function to isolate methyltransferases from an Escherichia coli whole cell lysate and identify them by MS.

Tags

MethyltransferasesS-adenosyl-L-homocysteine-binding ProteinsCapture Compound Mass SpectrometryAffinity ChromatographyActivity-based Protein ProfilingTrifunctional Capture CompoundsPhoto-crosslinkingSmall Molecule-protein InteractionsSelectivity FunctionSAHSorting FunctionBiotinSolid PhaseStreptavidin Magnetic BeadsOff-bead ConfigurationOn-bead ConfigurationSmall Molecule Of InterestSAMSAM-dependent MethyltransferasesDNA Methylation

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