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Abstract No.: ThP-181
Session: Peptide Sequencing
Presentation date: Thu, Aug 31, 2006
Presentation time: 09:50 – 11:20

An Enhanced Method for Peptides Sequencing Utilizing Stable Isotope Labeling

Marek Jan Noga1, Arndt Asperger2, Jerzy Silberring1

1 Jagiellonian University, Krakow, Poland
2 Bruker Daltonics, Bremen, Germany

Correspondence address: Marek Jan Noga, Jagiellonian University, Department of Neurobiochemistry, Ingardena 3, Krakow, 30-060 Poland.

Keywords: Labeling, Isotope; MS/MS; Protein Sequencing; Proteomic.

Novel aspect: High throughtput unambigous peptides sequencing by tandem mass spectrometry.

 

Identification of new protein and unknown protein modifications requires sequencing of minute amounts of peptides. De novo sequencing by means of tandem mass spectrometry is often a difficult and complex task due to unambiguosity of the acquired MS/MS spectra. In the method described here, stable isotope labeling allows for significant simplification of the data set. An isotopic label is atached to peptide’s N-terminus by derivatization with 1:1 mixture of acetic anhydride and deuterated acetic anhydride. During fragmentation all N-terminal fragments are labeled, allowing for highly sensitive, fast and reliable determination of peptide sequences. The method was tested in conjuction with both nano-ESI-MS/MS and MALDI-TOF/TOF approaches by successful sequencing of several standard peptide maps. This approach was applied for sequencing and identification of proteins from rat hipocampus with help of nano-scale two dimensional liquid chromatography (2D nano-LC-MS/MS). The technique is also suitable for quantitative measurements of proteins derived from biological samples.

Marek Noga is supported by EU and Polish Government throught Academic Innovation for Malopolska Programme.