NR AXXS
AU Thalassinos,K.; Hilton,G.R.; Slade,S.E.; Sanghera,N.; Robinson,P.J.; Pinheiro,T.J.T.; Bowers,M.T.; Scrivens,J.H.
TI Use of a Travelling Wave Based Ion Mobility Mass Spectrometry Approach to Resolve Prion Proteins of Varying Conformations
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: P01.30
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Protein misfolding diseases have been proposed to be initiated by a change in protein shape. A new mass spectrometry technique (termed ion mobility) has recently been developed that is able to yield information regarding molecular shape. This may be able to provide an insight into the conformational changes of prion proteins by measuring the change in cross section of the protein on misfolding. Measurement of misfolded protein would aid diagnosis and promote understanding of the disease propagation.
Ion mobility mass spectrometry measures how quickly a gaseous ion moves through a drift cell that is filled with a buffer gas under high pressure with the influence of a weak electric field. More compact ions with a small collision cross section will drift more quickly than extended ions. A travelling wave based ion mobility mass spectrometry approach is utilised to measure the change in cross section of the prion protein on misfolding.
All experiments were carried out on a Synapt HDMS System (Waters, Manchester, UK) which has a hybrid quadrupole/ion mobility/orthogonal acceleration time-of-flight (oa-TOF) geometry. Ions are accumulated in a trap travelling wave (T-Wave) device and periodically released into the ion mobility separation (IMS) T-Wave where they separate according to their mobility. The ions are then propelled through a transfer T-Wave to the oa-TOF for mass analysis. Ion arrival time distributions are recorded by synchronization of the ao-TOF with the gated release of ions into the IMS T-Wave.
Recombinant prion samples of Syrian hamster prion protein, SHaPrP(23-231) and SHaPrP(90-231), were expressed in both predominately alpha-helix and misfolded predominately beta-sheet forms. Circular dichroism experiments were carried out in order to confirm the nature of the secondary structure of the prion proteins. Estimations of the rotationally averaged cross sections of the proteins were made by reference to standards of known cross section. The ability of the approach to differentiate the normal cellular prion protein from the experimentally misfolded prion protein is demonstrated.
AD K. Thalassinos, G. Hilton, S. Slade, N. Sanghera, P. Robinson, T. Pinheiro, J. Scrivens, University of Warwick, Biological Sciences, UK; M. Bowers, University of California Santa Barbara, Chemistry, USA
SP englisch
PO Schottland
EA pdf-Datei und Poster (Autorenliste um die Autorin M. Grabenauer ergänzt)