N-Terminal protein sequencing

Use a combination of chemistry and mass spectrometry to analyse the N-Terminus of one or many proteins in your sample. 

Protein Pilot settings for N-Terminal labelling

Why study the N-Terminal of a protein?

Many biological processes require post-translational modifications to occur on a protein. For example, the majority of peptides hormones are produced as pro-hormones that need to be proteolytically cleaved to be active. Therefore, workflows that enable the identification of the N-Terminal of one or many proteins can become very interesting. Using a combination of chemical modifications, enrichment methods and mass spectrometry, we can produce a map of the "N-Terminome" of a sample. This workflow can be used to study the N-Terminus of pure proteins, or to identify variations in the N-Termini of every detectable protein in a sample.

General workflow

  1. In N-Terminal mapping workflows, native proteins N-Termini are first protected with a chemical group, generally using amine dimethylation.
  2. Proteins are digested with a protease, which generates non-native N-Temini.
  3. The newly generated non-native N-Termini are labeled with a "targetable" chemical group.
  4. Using a strategy that targets the second labeling reaction, the samples are depleted from every peptide whose N-Terminus was not protected with the first amine dimethylation reaction.
  5. The result of this depletion will be a sample in which the only remaining peptides will be the ones who were protected with the first chemical reaction, which are the N-Termini peptides of every protein in the sample. These peptides will be analyzed by LC-MS/MS.

Data report

For N-Terminal sequencing of a pure protein

Data reports for N-Terminal sequencing include the MS and MS/MS spectra of the detected peptide and any additional information that may be needed.

Proteome wide N-Terminal sequencing

List of every detected and confirmed N-Terminal peptide. Any specific information that the project requires will also be included. 

Contact us to discuss your N-terminal protein sequencing experiment

My personal experience of collaborating with PhenoSwitch Bioscience was really amazing and great. Ribosomal protein paralogs have very limited number of amino acid differences. If it were not for PhenoSwitch, it would not have been possible to get such a high quality identification and quantification of duplicated ribosomal proteins in yeast. I am highly indebted and thankful for their perseverance, commitment, help and guidance during the tough times of optimization. If you you need to use MS for identification and quantification of your molecules, I strongly recommend you to talk to these guys!
Mustafa Malik Ghulam, Post Doctoral Fellow, RNA Therapeutic Institute, Umass Medical School, Worcester, Massachussetts

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Stephen Naylor, Ph.D., CEO at ReNeuroGen LCC, Milwaukee, USA
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Klaus Klarskov, Ph.D., Professor at Université de Sherbrooke, Canada
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