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Impact of deglycosylation methods on two‐dimensional gel electrophoresis and matrix assisted laser desorption/ionization‐time of flight‐mass spectrometry for proteomic analysis
Author(s) -
Fryksdale Beth G.,
Jedrzejewski Paul T.,
Wong David L.,
Gaertner Alfred L.,
Miller Brian S.
Publication year - 2002
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200207)23:14<2184::aid-elps2184>3.0.co;2-1
Subject(s) - chemistry , mass spectrometry , proteome , chromatography , matrix assisted laser desorption/ionization , glycosylation , time of flight mass spectrometry , two dimensional gel electrophoresis , proteomics , protein mass spectrometry , gel electrophoresis , trichoderma reesei , peptide mass fingerprinting , sample preparation , surface enhanced laser desorption/ionization , sample preparation in mass spectrometry , biochemistry , tandem mass spectrometry , desorption , enzyme , electrospray ionization , ionization , cellulase , ion , organic chemistry , adsorption , gene
Glycosylation is a common post‐translational modification that can add complexity to the proteome of many cell types. We used enzymatic and chemical methods of deglycosylation to treat a heavily glycosylated exoproteome sample from the filamentous fungus Trichoderma reesei. Deglycosylated samples were resolved on one‐dimensional (1‐D) and two‐dimensional (2‐D) gels in order to determine the effect of deglycosylation on the electrophoresis patterns and on the ability to identify proteins by peptide mass matching using matrix assisted laser desorption/ionization‐time of flight‐mass spectrometry (MALDI‐TOF‐MS) analysis of in‐gel tryptic digests. We found that deglycosylation of the protein sample resulted in different protein patterns on 1‐D and 2‐D gels, reduced the complexity of gel patterns, and enhanced the protein identification of some proteins via MALDI‐TOF‐MS. Deglycosylation with trifluoromethanesulfonic acid (TFMS) was found to be more effective than enzymatic treatments. These deglycosylation techniques may be employed in whole proteome analysis to locate glycosylated proteins and assist in their identification by MS.