From 15N metabolic labelling to targeted LC-MS; different strategies applied to Systems Biology

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SEMINAR at 1pm Wednesday 25th November 2009

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MIB LECTURE THEATRE

Speaker: DR THIERRY LE BIHAN Senior Research Fellow

Centre for System Biology at Edinburgh (CSBE), University of Edinburgh


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Abstract

This presentation is divided into 2 different topics:

  • An approach based on 15N metabolic labelling has been developed and used to characterize the proteome of the unicellular algae Ostreococcus tauri under various conditions (different times of the day and different nutrient conditions). Some issues, strength and weakness of the technique will be also presented.
  • In this study, an assay based on the use of a linear ion trap has been developed in order to detect and quantify low intensity peptides.

Absolute quantitative measurements of a few anchor peptides performed with AQUA peptides are combined with a more global relative quantitation based on a peptide counts approach (emPAI). Finally an emphasis is done between peptide identification (using Mascot for example) versus peptide detection. In this study, peptide detection is performed by comparing experimental versus calculated m/z ratio, retention time and specific fragmentation pattern.

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