Spojení kapilární elektroforézy s molekulovou a prvkovou desorpční hmotnostní spektrometrií pro analýzu metaloproteinů
| Title in English | Off-line coupling of capillary electrophoresis to molecular and element desorption mass spectrometry for metalloprotein analysis |
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| Authors | |
| Year of publication | 2013 |
| Type | Conference abstract |
| MU Faculty or unit | |
| Citation | |
| Description | The most common methods for analysis of metal-protein complexes are based on on-line coupling of column separation techniques to electrospray mass spectrometry (MS) and nebulizer-inductively coupled plasma MS. The alternative tools for the metalloprotein analysis are methods based on laser desorption: matrix-assisted laser desorption/ionization (MALDI) MS and substrate-assisted laser desorption inductively-coupled plasma (SALD ICP) MS. Here, a new approach for an off-line multidetection platform for metalloprotein/metallopeptide analysis of fractions collected from a single CE run was presented. CE fractions are collected on a special target and analyzed consecutively with MALDI MS and SALD ICP MS. The concept is demonstrated on the CE-MALDI/SALD ICP MS analysis of rabbit liver metallothionein (MT) isoforms. Fractions from CE were collected on polyethyleneglycol target with a thin golden layer in a subatmospheric deposition chamber. After matrix addition, MALDI mass spectra of MT apoforms were recorded from the fractions. Finally, SALD ICP provided information on Cd content in the fractions. Alternatively, MALDI mass spectra of undissociated MT complexes from selected fractions may be recorded using a colder MALDI matrix. In summary, it was shown that a single CE separation yielded both proteomic and metallomic information. The off-line approach is a viable alternative to on-line coupling with electrospray/nebulizer ICP MS and it does not require both mass spectrometers physically close to CE. In addition, the fractions may be archived and subjected to laser-induced fluorescence detection or on target digestion. |
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