Reference : MALDI in-source decay of high mass protein isoforms: application to alpha- and beta-t...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/2268/126806
MALDI in-source decay of high mass protein isoforms: application to alpha- and beta-tubulin variants.
English
Calligaris, David [Université de Liège - ULg > Département de chimie (sciences) > GIGA-R : Laboratoire de spectrométrie de masse (L.S.M.) >]
Villard, Claude [> >]
Terras, Lionel [> >]
Braguer, Diane [> >]
Verdier-Pinard, Pascal [> >]
Lafitte, Daniel [> >]
Jul-2010
Analytical Chemistry
American Chemical Society
82
14
6176-6184
Yes (verified by ORBi)
International
0003-2700
1520-6882
Washington
DC
[en] Tubulin is one of the major targets in cancer chemotherapy and the target of more than twenty percent of the cancer chemotherapic agents. The modulation of isoform content has been hypothesized as being a cause of resistance to treatment. Isoform differences lie mostly in the C-terminus part of the protein. Extensive characterization of this polypeptide region is therefore of critical importance. MALDI-TOF fragmentation of tubulin C-terminal domains was tested using synthetic peptides. Then, isotypes from HeLa cells were successfully characterized for the first time by in-source decay (ISD) fragmentation of their C-terminus coupled to a pseudo MS3 technique named T3-sequencing. The fragmentation occurred in-source, preferentially generating yn-series ions. This approach required guanidination for the characterization of the βIII-tubulin C-terminus peptide. This study is, to our knowledge, the first example of reflectron in-source decay (reISD) of the C-terminus of a 50 kDa protein. This potentially occurs via a CID-like mechanism occurring in the MALDI plume. There are now new avenues for top-down characterization of important clinical biomarkers such as βIII-tubulin isotypes, a potential marker of drug resistance and tumor progression. This paper raises the challenge of protein isotypes characterization for early cancer detection and treatment monitoring.
Researchers ; Professionals ; Students ; General public
http://hdl.handle.net/2268/126806
10.1021/ac100996v
http://pubs.acs.org/doi/abs/10.1021/ac100996v

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