Nano Archive

Characterising electrosprayed biomolecules using tandem-MS--the noncovalent GroEL chaperonin assembly

Sobott, Frank and Robinson, Carol V. (2004) Characterising electrosprayed biomolecules using tandem-MS--the noncovalent GroEL chaperonin assembly. International Journal of Mass Spectrometry, 236 (1-3). 25 - 32.

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It is now possible to transfer large protein complexes intact into the gas phase using nano-electrospray ionisation (ESI) and to investigate their stoichiometry in a mass spectrometer. Using as a model assembly the previous termnoncovalentnext term 14mer (ca. 800 kDa) of the chaperonin GroEL from Escherichia coli we show that the measured mass of the assembly is higher than expected from the sum of the components and explore parameters of ion activation that affect this ‘previous termnoncovalentnext term mass shift’. Under optimal desolvation conditions the measured mass is not, vert, similar0.5% greater than the calculated value indicating that part of the solution environment remains attached to these ions during phase transfer. The origin of this previous termnoncovalentnext term mass shift is explored using previous termtandemnext term mass spectrometry experiments. Collisional activation of the 65+ charge state of the GroEL 14mer indicates the presence of up to 100 solvent/buffer molecules, both positively and negatively charged, which are stripped during CID in the gas-filled collision cell. At high collision energies, asymmetric dissociation into highly charged monomer and 13mer complexes takes place revealing the subunit composition of the assembly. Under these conditions the previous termnoncovalentnext term mass shift is significantly reduced (<0.04%) demonstrating the utility of this previous termtandemnext term approach for mass measurement of previous termbiomoleculesnext term in the gas phase.

Item Type:Article
Additional Information:Special issue: In honour of Dudley H. Williams
Uncontrolled Keywords:Nano-electrospray; previous termTandemnext term-mass spectrometry; Collision-induced dissociation; previous termNoncovalentnext term interactions; Protein complexes
Subjects:Biomedical Science > Nanomedicine
ID Code:5789
Deposited By:SPI
Deposited On:23 Jul 2009 16:04
Last Modified:23 Jul 2009 16:04

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