Mass Spectrometry and Proteomics

Keywords: mass spectrometry, MS, MALDI, nanoLC-MS/MS, proteomics, 2-D gel electrophoresis

The Mass Spectrometry and Proteomics platform provides a state-of-the-art proteomics service for researchers at the Institut Curie, as well as for collaborators from external national and international institutions, to identify and characterise proteins and their post-translational modifications. In addition, the platform offers workshops on preparing biological samples for mass spectrometry and separation of proteins by two-dimensional gel electrophoresis.

The platform is located in the Developmental Biology and Cancer building on the central Paris site. It has three mass spectrometers:

The two ESI tandem mass spectrometers are able to gives information about peptide sequences.

A hybrid Fourier Transform Mass Spectrometer (Thermo Scientific LTQ Orbitrap XL), supporting a wide range of applications from routine compound identification to the most challenging analysis of low level components in complex mixtures.

A QUAD-TOF (Applied Biosystems Qstar® Elite), which gives better information about peptide sequences. This technique is useful for solving ambiguities about peptide identity and/or for de novo protein sequencing when little or nothing is known about the genome sequence. To investigate complex mixtures of peptides, the mass spectrometers are coupled to a high-performance liquid chromatography system (two Dionex U3000).

Our MALDI TOF-TOF tandem mass spectrometer (Applied Biosystems 4800 plusTM) is used to identify, by high-resolution peptide mass fingerprinting, proteins from species whose genomes have been sequenced. The use of tandem time-of-flight MS/MS provides highest level of protein coverage, throughput, and confidence in quantitative proteomic analysis. The analyzer combines all advantages of MALDI and is working in complementary fashion to the LTQ Orbitrap XL and/or Qstar Elite.

The mass spectrometry data are submitted to two different but complementary algorithms to obtain the corresponding protein names from the databases. The data so obtained are entered into our reference database (myProMS) to allow efficient data management, curation and sharing between mass spectrometrists and biologists.

The service participates in the Cancéropôle Ile-de-France and, in this context, it collaborates especially with the Gustave-Roussy Institute (Villejuif) and the École supérieure de physique et de chimie industrielles (ESPCI, Paris). Our main collaborations, however, are with groups from the research section at the Institut Curie. We have on-going long-term projects on the proteomics of phagosomes in dendritic cells and the proteomics of exosomes (with researchers in the Immunity and Cancer unit, Unit 653 Inserm/Institut Curie), as well as on the use of toxins as vectors for anti-tumour drugs (with researchers in the Subcellular Structure and Cellular Dynamics unit, UMR144 CNRS/Institut Curie). In addition, we work with groups interested in development biology (at the neighbouring École normale supérieure) and in parasitology (at the Pasteur Institute in Paris and at New York University, USA).

Last update: March 2009