Study of ionization of tryptic peptides on electrospray ionization mass spectrometry
2005
Araki, T. (Kyushu Tokai Univ., Choyo, Kumamoto (Japan). Faculty of Agriculture) | Shigetome, R. | Takaira, A. | Torikata, T.
In the recent proteomics study, 2D-PAGE is the most effective method for separation of proteins followed by mass spectrometry analysis. However, the relation of treatment of peptide samples and peptide properties are not elucidated. In the present study, we investigated about the conditions for peptide sequencing using quadrupole tune of fright mass spectrometry analysis. Namely, the condition of ingel digestion of protein spot from SDS-PAGE and concentration of enzyme was estimated. Further, the relation of peptide properties and recovery of peptide ions on mass spectrometry was analyzed. The most effective concentration of trypsin was found to be 0.01 micro g/ micro l. For the pretreatment of proteins before ingel digestion, the reductive alkylation without denaturant was found to be effective. The peptide ionization on mass spectrometry depended on the property of apparatus that showed the higher recovery of peptide ion in the region of m/z of 300 to 800. The ionization of peptide was slightly depended on the hydrophobicity of peptide and showed the lower recovery for extra hydrophobic or hydrophilic peptides. ZipTip treatment, for peptide purification, showed the effective for peptides found in the region of m/z of 400 to 800, but the loss of recovery outside of that region.
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