Detection of the intact GroEL chaperonin assembly by mass spectrometry

Journal of the American Chemical Society

Volumn 121, Issue 19, 1999, Pages 4718-4719

  Rostom, Adam A ^a   Robinson, Carol V ^a  

^a UNIVERSITY OF OXFORD   (United Kingdom)

Author keywords

[No Author keywords available]

Indexed keywords

CHAPERONIN;

ARTICLE; ENTROPY; HYDROPHOBICITY; MACROMOLECULE; MASS SPECTROMETRY; MOLECULAR WEIGHT; PROTEIN ASSEMBLY; PROTEIN DETERMINATION; PROTEIN FOLDING; STOICHIOMETRY;

EID: 0033583731     PISSN: 00027863     EISSN: None     Source Type: Journal    
DOI: 10.1021/ja990238r     Document Type: Article

References (13)

1. 12 a quadruple mass analyser, followed by a hexapole collision cell and a time-of-flight mass analyser. Ions are focused by an rF lens before transmission to the quadrupole which is used in rF-only mode as a wide band-pass filter. The ions are then conducted through the hexapole collision cell, pressurized with dry argon. Variation of the collision energy up to 140 V, induces dissociation of the macromolecular complex via collision with argon gas molecules. Ions are then pulsed into the ToF with an acceleration voltage of 8 kV at a pulse rate of 4 kHz for detection with a microchannel plate detector. Acquisition is made through a time-to-digital converter operating at 1 GHz. Mass spectra were obtained without source heating and in the absence of organic cosolvents. Mass spectrometer tuning parameters were held constant with a needle voltage of 1550 V, skimmer cone voltage of 150 V, skimmer offset of 4 V, and a collision energy of 4 V, except where stated.
2. 4 Pa backing pressure during the electrospray process.
3. Robinson, C. V.; Groß, M.; Eyles, S. J.; Ewbank, J. J.; Mayhew, M.; Hartl, F. U.; Dobson, C. M.; Radford, S. E. Nature 1994, 572, 646-651.
4. Gervasoni, P.; Staudenman, W.; James, P.; Gehrig, P.; Pluckthun, A. Proc. Natl. Acad. Sci. U.S.A. 1996, 93, 12189-12194.
5. Ayed, A.; Krutchinsky, A. N.; Ens, W.; Standing, K. G.; Duckworth, H. W. Rapid Commun. Mass Spectrom. 1998, 12, 339-344.
6. Rostom, A. A.; Sunde, M.; Richardson, S. J.; Schreiber, G.; Jarvis, S.; Bateman, R.; Dobson, C. M.; Robinson, C. V. Proteins: Struct., Funct., Genet. 1998, Suppl. 2, 3-11.
7. Nettleton, E. J.; Sunde, M.; Lai, V.; Kelly, J. W.; Dobson, C. M.; Robinson, C. V. J. Mol. Biol. 1998, 281, 553-564.
8. Braig, K.; Otwinowski, Z.; Hedge, R.; Boisvert, D. C.; Joachimiak, A.; Horwich, A. L.; Sigler, P. B. Nature 1994, 62, 349-357.
9. -5 Pa. Mass spectra were calibrated with CsI (Sigma Chemical Co.). Micromass Mass Lynx software version 3.1 was used for data processing.
10. Krutchinsky, A. N.; Chernushevich, I.; Spicer, V.; Ens, W.; Standing, K. G. J. Am. Soc. Mass Spectrom. 1998, 9, 569-579.
11. Verentchikov, A.; Ens, W.; Standing, K. Anal. Chem. 1994, 66, 126-133.
12. Wilm, M.; Mann, M. Int. J. Mass Spectrom. and Ion Processes 1994, 136, 167-180.
13. Todd, M.; Lorimer, G. Methods Enymol. 1998, 135-141.