Project leader: Professor Per O. Seglen

Project staff: Monica Fengsrud, dr. scient.; Hamid R. Samari, dr. scient.; Anders Øverbye, cand.scient.; Marianne Lunde Sneve, cand.scient.; Siri Hildonen, cand.scient.; Tonje Asmyhr, cand.scient.; Frank Sætre, cand.scient., dept.eng.; Linda Korseberg Hagen, dept.eng.

Web page: https://www.ous-research.no/seglen

Objectives

To offer the competence, instrumentation and methods needed to help scientists at DNR and RH in addressing the following issues:

• Analysis of large proteomes (tissue samples; subcellular fractions) by multidimensional chromatographic and electrophoretic fractionation, e.g., isoelectric batch focusing, ultrafiltration, affinity-, ion exchange or RP-LC chromatography, followed by one- or two-dimensional (2D) gel electrophoresis.
• Identification of individual proteins (e.g., from 2D-gels) by peptide mass fingerprinting, using MALDI-TOF mass spectrometry of tryptic peptides.
• Sequencing of unknown peptides by nanoelectrospray injection and ion trap mass spectrometry.
• Identification of protein phosphorylation sites by a combination of MALDI-TOF and ion trap mass spectrometry.

Furthermore, the Proteomics Section will work towards the development of proteomic methods suitable for differential cancer diagnosis, with emphasis on the following aspects:

• Separation of the diagnostically interesting low-abundance proteins in human serum from the trivial high-abundance serum proteins (albumin, immunoglobulins etc.).
• Multidimensional fractionation of serum by the separation methods outlined above.
• N-terminal protein tagging followed by tryptic proteolysis and chromatographic isolation of N-terminal peptides.
• Analysis (and, if needed, identification by sequencing) of the tryptic peptides by ion trap mass spectrometry.
• Bioinformatic processing (pattern analysis) of the peptide mass list to produce a meaningful differential diagnosis.

Milestones

• 1997-1998: Establishment of two-dimensional gel electrophoresis methods for protein separation at the Department of Cell Biology.
• 2001: Foundation of the Proteomics Section (Proteomics and Mammalian Cell Biology Section). Acquisition of a MALDI-TOF mass spectrometer.
• 2002: Publication of the first Norwegian paper describing an in-house protein identification by proteomic methods (Ruud Larsen et al., J. Biol. Chem. 277, 34826-34835).
• 2003: Protein identification by peptide mass fingerprinting offered as a routine service for scientists at the University of Oslo, with support from the EMBIO (UiO) program.
• 2005: Acquisition of an ion trap mass spectrometer and chromatographic equipment for nanoelectrospray injection.