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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Bertoletti, Laura Regazzoni, Luca Aldini, Giancarlo Colombo, Raffaella Abballe, Franco Caccialanza, Gabriele De Lorenzi, Ersilia |
| Description | Country affiliation: Italy Author Affiliation: Bertoletti L ( Department of Drug Sciences, University of Pavia, Pavia, Italy. laura.bertoletti@unipv.it) |
| Abstract | In this work we present for the first time the use of ion-exchange liquid chromatography to separate the native form and a partially structured intermediate of the folding of the amyloidogenic protein beta2-microglobulin. Using a strong anion-exchange column that accounts for the differences in charge exposure of the two conformers, a LC-UV method is initially optimised in terms of mobile phase pH, composition and temperature. The preferred mobile phase conditions that afford useful information were found to be 35 mM ammonium formate, pH 7.4 at 25°C. The dynamic equilibrium of the two species is demonstrated upon increasing the concentration of acetonitrile in the protein sample. Then, the chromatographic method is transferred to MS detection and the respective charge state distributions of the separated conformers are identified. The LC-MS results demonstrate that one of the conformers is partially unfolded, compared with the native and more compact species. The correspondence with previous results obtained in free solution by capillary electrophoresis suggest that strong ion exchange LC-MS does not alter beta2-microglobulin conformation and maintains the dynamic equilibrium already observed between the native protein and its folding intermediate. |
| File Format | HTM / HTML |
| ISSN | 00032670 |
| Volume Number | 771 |
| e-ISSN | 18734324 |
| Journal | Analytica Chimica Acta |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2013-04-10 |
| Publisher Place | Netherlands |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Analytical Discipline Chemistry Chromatography, Ion Exchange Methods Mass Spectrometry Protein Folding Beta 2-microglobulin Chemistry Isolation & Purification Protein Conformation Spectrometry, Mass, Electrospray Ionization Journal Article Research Support, Non-u.s. Gov't |
| Content Type | Text |
| Resource Type | Article |
| Subject | Spectroscopy Environmental Chemistry Analytical Chemistry Biochemistry |
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