Institut de Biologie StructuraleGrenoble / France

Contact person(s) related to this article / BOERI-ERBA Elisabetta



Head of platform : Elisabetta Boeri Erba

We are interested in studying protein complexes by native mass spectrometry (native MS). This specific MS technique allows the preservation within the gas phase of non-covalent interactions between subunits of a protein assembly. After different steps of analysis (Figure 1) one can build 2D interaction maps of protein complexes. Native MS represents a powerful and appropriate tool to study the architecture of protein complexes of different sizes (from some tens of kDa up to MDa) when high-resolution structural data are not available. It complements the existing methods in structural biology since it requires very little amounts of sample (5 to 10 μM of a protein complex are sufficient for a successful analysis), tolerates heterogeneity of subunit composition, does not require crystals, and symmetry in the structure does not represent an advantage

Fig 1. A 2D interaction network of subunits within protein complexes can be generated through a multi-step process using native mass spectrometry (native MS).
Step 1. Under denaturing conditions, the subunits of a complex are chromatographically separated and their masses are determined.
Step 2. Using MS conditions optimized for preserving non-covalent interactions, the measured mass of the intact complex reveals the stoichiometry of the subunits.
Step 3. A series of tandem MS spectra indicates which are subunits located within the core and which are peripheral proteins.
Step 4. By adding organic solvent, overlapping sub-complexes (e.g. dimers, trimers) are generated. The composition of the different sub-complexes reveals the interactions between the subunits.
Step 5. Individual subunits can be mixed in solution and a mass shift can be detected if a subcomplex is generated.
Step 6. The combination of all these data allows one to draw an accurate 2D interaction network of a protein complex.

Key Words :

Native mass spectrometry, protein complexes, non-covalent interactions, 2D interaction maps.

Techniques :

- Expression and purification of protein complexes
- Native mass spectrometry
- Protein biophysical characterization

Platform members :

- Elisabetta Boeri Erba, CEA Research Engineer
- Luca Signor, CEA Research Engineer

Available services :

- Mass spectrometry platform

Publications :

Boeri Erba E, Signor L, Oliva MF, Hans F, Petosa C. Characterizing Intact Macromolecular Complexes Using Native Mass Spectrometry. Methods Mol Biol. 2018 ;1764:133-151.

Alfieri A, Sorokina O, Adrait A, Angelini C, Russo I, Morellato A, Matteoli M, Menna E, Boeri Erba E, McLean C, Armstrong JD, Ala U, Buxbaum JD, Brusco A, Couté Y, De Rubeis S, Turco E, Defilippi P. Synaptic Interactome Mining Reveals p140Cap as a New Hub for PSD Proteins Involved in Psychiatric and Neurological Disorders. Front Mol Neurosci. 2017 10:212.

Pellegrini E, Signor L, Singh S, Boeri Erba E, Cusack S. Structures of the inactive and active states of RIP2 kinase inform on the mechanism of activation. PLoS One. 2017 12(5):e0177161.

Macek P, Kerfah R, Boeri Erba E, Crublet E, Moriscot C, Schoehn G, Amero C, Boisbouvier J. Unraveling self-assembly pathways of the 468-kDa proteolytic machine TET2. Sci Adv. 2017 3(4):e1601601.

Sauer PV, Timm J, Liu D, Sitbon D, Boeri-Erba E, Velours C, Mücke N, Langowski J, Ochsenbein F, Almouzni G, Panne D. Insights into the molecular architecture and histone H3-H4 deposition mechanism of yeast Chromatin assembly factor 1. Elife. 2017 Mar 18 ;6. pii : e23474.

Lopez S, Rondot L, Cavazza C, Iannello M, Boeri-Erba E, Burzlaff N, Strinitz F, Jorge-Robin A, Marchi-Delapierre C, Ménage S. Efficient conversion of alkenes to chlorohydrins by a Ru-based artificial enzyme. Chem Commun. 2017 53:3579-3582.

Baquero E, Albertini AA, Raux H, Abou-Hamdan A, Boeri-Erba E, Ouldali M, Buonocore L, Rose JK, Lepault J, Bressanelli S, Gaudin Y. Structural intermediates in the fusion-associated transition of vesiculovirus glycoprotein. EMBO J. 2017 36:679-692.

Rasmussen KK, Frandsen KE, Boeri Erba E, Pedersen M, Varming AK, Hammer K, Kilstrup M, Thulstrup PW, Blackledge M, Jensen MR, Lo Leggio L. Structural and dynamics studies of a truncated variant of CI repressor from bacteriophage TP901-1. Sci Rep. 2016 6:29574.

Yee AW, Moulin M, Breteau N, Haertlein M, Mitchell EP, Cooper JB, Boeri Erba E, Forsyth VT. Impact of Deuteration on the Assembly Kinetics of Transthyretin Monitored by Native Mass Spectrometry and Implications for Amyloidoses. Angew Chem Int Ed Engl. 2016 55:9292-6..

Caillat C, Macheboeuf P, Wu Y, McCarthy AA, Boeri-Erba E, Effantin G, Göttlinger HG, Weissenhorn W, Renesto P. Asymmetric ring structure of Vps4 required for ESCRT-III disassembly. Nat Commun. 2015 6:8781.

Boeri Erba E, Petosa C (2015) The emerging role of native mass spectrometry in characterizing the structure and dynamics of macromolecular complexes. Protein Sci. 24(8):1176-92. doi : 10.1002/pro.2661

Boeri Erba E, Klein PA, Signor L. (2015) Combining a NHS ester and glutaraldehyde improves crosslinking prior to MALDI MS analysis of intact protein complexes. J Mass Spectrom. 50(10):1114-9. doi : 10.1002/jms.3626

Boeri Erba E (2014) Investigating macromolecular complexes using top-down mass spectrometry. Proteomics. 14(10):1259-70. doi : 10.1002.

Signor L, Boeri Erba E. (2013) Matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometric analysis of intact proteins larger than 100 kDa. J Vis Exp, (79). doi : 10.3791/50635.

Levy ED, Boeri Erba E, Robinson CV, Teichmann SA. (2008) Assembly reflects evolution of protein complexes. Nature. 453 : 1262-5.