remove the remaining detergent molecules. Simultaneously
and incrementally increasing the Trap gas flow (from 2 mL/
min up to 8 mL/min) might improve the transmission of these
particular large or membrane protein complexes.
Acknowledgments
The authors would like to thank Arnaud Poterszman (CAK complex), Albert Weixlbaumer (E. coli RNA polymerase), and Isabelle
Billas Massobrio (ERR complexes) from the IGBMC (Strasbourg,
France) and Virginie Gervais and Alain Milon (p8 protein) from
IPBS (Toulouse, France) for kindly providing samples. This work
was supported by the French Ministry of Research (Investissements
d’Avenir Program, Proteomics French Infrastructure, ANR-10INBS-08) and the FEDER (Fonds Europe ´ens de De ´veloppement
Re ´gional), Toulouse Me ´tropole, and the Re ´gion Midi-Pyre ´ne ´es.
References
1. Ganem B (1991) Detection of noncovalent
receptor-ligand complexes by mass spectrometry. J Am Chem Soc 113:6294–6296
2. Ganem B (1991) Observation of noncovalent
enzyme-substrate and enzyme-product complexes by ion-spray mass spectrometry. J Am
Chem Soc 113:7818–7819
3. Katta V, Chait B (1991) Observation of the
hemoglobin complex in native myoglobin by
electrospray-ionization mass spectrometry. J
Am Chem Soc 113:8534–8535
4. Nettleton EJ, Sunde M, Lai Z, Kelly JW, Dobson CM, Robinson CV (1998) Protein subunit
interactions and structural integrity of amyloidogenic transthyretins: evidence from electrospray mass spectrometry. J Mol Biol 281
(3):553–564
5. Pinkse MWH, Heck AJR, Rumpel K, Pullen F
(2004) Probing noncovalent protein-ligand
interactions of the cGMP-dependent protein
kinase using electrospray ionization time of
flight mass spectrometry. J Am Soc Mass Spectrom 15(10):1392–1399
6. Sobott F, Herna ´ndez H, McCammon MG,
Tito MA, Robinson CV (2002) A tandem
mass spectrometer for improved transmission
and analysis of large macromolecular assemblies. Anal Chem 74(6):1402–1407
7. Rose RJ, Damoc E, Denisov E, Makarov A,
Heck AJ (2012) High-sensitivity Orbitrap
mass analysis of intact macromolecular assemblies. Nat Methods 9(11):1084–1086
8. Andersson FI, Tryggvesson A, Sharon M, Diemand AV, Classen M, Best C, Schmidt R,
Schelin J, Stanne TM, Bukau B, Robinson
CV, Witt S, Mogk A, Clarke AK (2009) Structure and function of a novel type of
ATP-dependent Clp protease. J Biol Chem
284(20):13519–13532
9. Sharon M, Mao H, Boeri Erba E, Stephens E,
Zheng N, Robinson CV (2009) Symmetrical
modularity of the COP9 signalosome complex
suggests its multifunctionality. Structure 17
(1):31–40
10. Martinez-Zapien D, Saliou JM, Han X,
Atmanene C, Proux F, Cianfe ´rani S, DockBregeon AC (2015) Intermolecular recognition of the non-coding RNA 7SK and
HEXIM protein in perspective. Biochimie
117:63–71
11. Saliou JM, Manival X, Tillault AS,
Atmanene C, Bobo C, Branlant C, Van
Dorsselaer A, Charpentier B, Cianfe ´rani S
(2015) Combining native MS approaches to
decipher archaeal box H/ACA ribonucleoprotein particle structure and activity. Proteomics
15(16):2851–2861
12. Barrera NP, Di Bartolo N, Booth PJ, Robinson
CV (2008) Micelles protect membrane complexes from solution to vacuum. Science 321
(5886):243–246
13. Chorev DS, Baker LA, Wu D, BeilstenEdmands
V,
Rouse
SL,
Zeev-BenMordehai T, Jiko C, Samsudin F, Gerle C,
190
Ste ´ phane Erb et al.
and incrementally increasing the Trap gas flow (from 2 mL/
min up to 8 mL/min) might improve the transmission of these
particular large or membrane protein complexes.
Acknowledgments
The authors would like to thank Arnaud Poterszman (CAK complex), Albert Weixlbaumer (E. coli RNA polymerase), and Isabelle
Billas Massobrio (ERR complexes) from the IGBMC (Strasbourg,
France) and Virginie Gervais and Alain Milon (p8 protein) from
IPBS (Toulouse, France) for kindly providing samples. This work
was supported by the French Ministry of Research (Investissements
d’Avenir Program, Proteomics French Infrastructure, ANR-10INBS-08) and the FEDER (Fonds Europe ´ens de De ´veloppement
Re ´gional), Toulouse Me ´tropole, and the Re ´gion Midi-Pyre ´ne ´es.
References
1. Ganem B (1991) Detection of noncovalent
receptor-ligand complexes by mass spectrometry. J Am Chem Soc 113:6294–6296
2. Ganem B (1991) Observation of noncovalent
enzyme-substrate and enzyme-product complexes by ion-spray mass spectrometry. J Am
Chem Soc 113:7818–7819
3. Katta V, Chait B (1991) Observation of the
hemoglobin complex in native myoglobin by
electrospray-ionization mass spectrometry. J
Am Chem Soc 113:8534–8535
4. Nettleton EJ, Sunde M, Lai Z, Kelly JW, Dobson CM, Robinson CV (1998) Protein subunit
interactions and structural integrity of amyloidogenic transthyretins: evidence from electrospray mass spectrometry. J Mol Biol 281
(3):553–564
5. Pinkse MWH, Heck AJR, Rumpel K, Pullen F
(2004) Probing noncovalent protein-ligand
interactions of the cGMP-dependent protein
kinase using electrospray ionization time of
flight mass spectrometry. J Am Soc Mass Spectrom 15(10):1392–1399
6. Sobott F, Herna ´ndez H, McCammon MG,
Tito MA, Robinson CV (2002) A tandem
mass spectrometer for improved transmission
and analysis of large macromolecular assemblies. Anal Chem 74(6):1402–1407
7. Rose RJ, Damoc E, Denisov E, Makarov A,
Heck AJ (2012) High-sensitivity Orbitrap
mass analysis of intact macromolecular assemblies. Nat Methods 9(11):1084–1086
8. Andersson FI, Tryggvesson A, Sharon M, Diemand AV, Classen M, Best C, Schmidt R,
Schelin J, Stanne TM, Bukau B, Robinson
CV, Witt S, Mogk A, Clarke AK (2009) Structure and function of a novel type of
ATP-dependent Clp protease. J Biol Chem
284(20):13519–13532
9. Sharon M, Mao H, Boeri Erba E, Stephens E,
Zheng N, Robinson CV (2009) Symmetrical
modularity of the COP9 signalosome complex
suggests its multifunctionality. Structure 17
(1):31–40
10. Martinez-Zapien D, Saliou JM, Han X,
Atmanene C, Proux F, Cianfe ´rani S, DockBregeon AC (2015) Intermolecular recognition of the non-coding RNA 7SK and
HEXIM protein in perspective. Biochimie
117:63–71
11. Saliou JM, Manival X, Tillault AS,
Atmanene C, Bobo C, Branlant C, Van
Dorsselaer A, Charpentier B, Cianfe ´rani S
(2015) Combining native MS approaches to
decipher archaeal box H/ACA ribonucleoprotein particle structure and activity. Proteomics
15(16):2851–2861
12. Barrera NP, Di Bartolo N, Booth PJ, Robinson
CV (2008) Micelles protect membrane complexes from solution to vacuum. Science 321
(5886):243–246
13. Chorev DS, Baker LA, Wu D, BeilstenEdmands
V,
Rouse
SL,
Zeev-BenMordehai T, Jiko C, Samsudin F, Gerle C,
190
Ste ´ phane Erb et al.