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Integrated Structural Biology Grenoble
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Contacts relatifs à cet article / SIGNOR Luca

Mass spectrometry

Presentation

The mass spectrometry platform at IBS provides the scientific community a service for protein analysis and characterization and offers the following analytical techniques :

  • Determination of exact mass, at high resolution, for intact proteins and peptides
  • Control of protein and peptide expression, modification, mutation and labeling
  • Analyses of protein limited proteolysis
  • Characterization of purified proteins after enzymatic digestion (in-gel or in solution)
  • Analysis of protein complexes by native mass spectrometry.

The mass spectrometry platform has been engaged in developing quality management which led in June 2011 to ISO 9001 (version 2008) certification. The quality management allows an optimal follow up of competences and equipment, of scientific information (traceability of data, analyses and projects), better visibility and transparency of used procedures and a continuous adequacy with the needs expressed by the users.

The activities proposed by the mass spectrometry platform can be carried in two ways :

Routine analyses : corresponding to priced analyses which do not need of a particular technical development (Quality Control)

Collaborative projects : involving participation in a research project or collaboration, the development of a specific method and/or a technological development (Research and Developpement)

The methods proposed in the context of quality control are detailed in the document attached herewith :

Analyses methods

Development and application of other methods of analysis by mass spectrometry will be possible after prior discussion with the staff of the platform.

Key words

Mass Spectrometry (MS), Electrospray Ionization (ESI), MALDI, Time-of-Flight (TOF), High Resolution, Liquid Chromatography (LC), Protein and Peptide analyses, Limited proteolysis, Native Mass Spectrometry (Native MS).

Dedicated Staff

Elisabetta Boeri Erba, CEA researcher-engineer.

Luca Signor, CEA researcher-engineer.

Equipement

MALDI-TOF MS (Autoflex, Bruker Daltonics)
ion source : MALDI
analyser : TOF
m/z up to 500’000 (linear mode)
resolution : up to 20’000 (reflectron mode).

LC ESI-TOF MS (6210, Agilent Technologies)
ion source : ESI et nanoESI (HPLC-Chip)
analyser : TOF
m/z 30-3000
resolution : > 20’000
coupled to capillary and nano HPLC system (séries 1100).

High mass ESI Q-TOF MS (Ultima, Micromass Waters)
ion source : ESI et nanoESI
analyser : TOF
m/z 30-32000
resolution : > 10’000
coupled to capillary and nano HPLC system (CapLC Waters).

Instrument monitoring : the spectrometers are checked up regularly (preventive and corrective maintenance service contract) and are calibrated before each series of measurements.

Accessibility

IBS, PSB members, external users (academia, industry).

How to make a request ?

1. Before submitting the analysis request form and if you have any question concerning sample preparation and the choice of the more appropriate analysis method, we suggest you contacting us for discussion.

2. Fill out the analysis request form (AR (save the empty file before filling it) and send it by e-mail to : ibs-plateforme-ms.contact(at)ibs.fr.

Analysis request

The analysis request form is validated at sample reception by the staff of the platform.

3. The samples are collected at IBS, room 166 ou 167 (monday-friday, 9h30-17h30).

4. The analysis will start only after validation of the analysis request form by the user (signing of the AR form by the user at sample reception).

Contact et localisation


ibs-plateforme-ms.contact(at)ibs.fr

IBS
Office : room 166 / 167
Laboratory : rooms 159 / 159A

Samples

Toxic and/or radioactive samples are not accepted. Only the biological samples whose biohazard level is 1 (non-pathogenic) will be accepted.

A unique analysis number is assigned to each analysis request for sample identification. Sample tubes must be clearly annotated with an ID corresponding to that reported on the AR form (refer to sample list).

Samples will be stored at room temperature or 4 ° C or -20 ° C (depending on the choice indicated on the AR form). The samples will not be stored after analysis unless specified by the client.

Optimal conditions for MS analysis :

  • Use water of high quality for all solutions (HPLC or MilliQ quality).
  • Buffers and additives : avoid phosphate (PBS) and sulphate buffers, detergents (e.g. Triton, CHAPS...) and contamination by polymers (e.g. polyethylene glycols).
  • Sample concentration and volume : ≥ 10 microM and minimum volume 20 µl
  • For samples into solution is recommended filtering or centrifuging prior to analysis in order to eliminate any insoluble substance and suspended particles.

Results

Results are sent by e-mail as pdf format (analysis report). In general delay time is 48 hours after sample reception (for standard analyses, limited number and good quality samples) ; a more precise delay time will be defined with the user on an individual basis upon sample receipt. Certificates of analysis and raw data will be stored and available for 5 years.

Cost

Analyses prices are defined and adapted depending on the type of service and project requested by the customer. Contact us for a quote.

Follow up / Development

Routine analyses : the user agrees to promote the obtained results by mentioning the platform in the acknowledgements in case of publication or scientific communication.

In the general case users must aknowledge the platform in their publications by using the sentence :
« This work used the platforms of the Grenoble Instruct-ERIC center (ISBG ; UMS 3518 CNRS-CEA-UGA-EMBL) within the Grenoble Partnership for Structural Biology (PSB), supported by FRISBI (ANR-10-INBS-05-02) and GRAL, financed within the University Grenoble Alpes graduate school (Ecoles Universitaires de Recherche) CBH-EUR-GS (ANR-17-EURE-0003) ».

Collaborative projects : collaborative projects imply that the platform staff that have been actively involved in the project will be co-authors of the published papers or communications.

Selected publications

Zapun A, Philippe J, Abrahams K, Signor L, Roper D, Breukink E, Vernet T.
In vitro reconstitution of peptidoglycan assembly from the Gram-positive pathogen Streptococcus pneumoniae.
ACS Chemical Biology, 2013, 8 (12), 2688–2696.

Noirclerc-Savoye M, Lantez V, Signor L, Philippe J, Vernet T, Zapun A.
Reconstitution of membrane protein complexes involved in pneumococcal septal cell wall assembly.
PLoS ONE, 2013, 8(9), e75522.

Parent A, Caux-Thang C, Signor L, Clémancey M, Sethu R, Blondin G, Maldivi P, Duarte V, Latour J-M.
A single aspartate to glutamate mutation makes Fur sensitive to H2O2 as PerR.
Angewandte Chemie Int. Ed., 2013, 52, 10339-10343.

Dach I, Olesen C, Signor L, Nissen P, le Maire M, Møller JV, Ebel C.
Active detergent solubilized H+,K+-ATPase is a monomer.
Journal of Biological Chemistry, 2012, 287(50), 41963-41978.

Wollers S, Layer G, Garcia-Serres R, Signor L, Clemancey M, Latour JM, Fontecave M, Ollagnier de Choudens S.
Iron-sulfur (Fe-S) cluster assembly : the SufBCD complex is a new type of Fe-S scaffold with a flavin redox cofactor.
Journal of Biological Chemistry, 2010, 285(30), 23331-23341.