Label-Free Analysis of O-glycosylation Site-Occupancy Based on the Signal Intensity of Glycopeptide/Peptide Ions

• Wada Yoshinao

  Department of Molecular Medicine, Osaka Medical Center and Research Institute for Maternal and Child Health

抄録

Mucin-type O-glycosylation is a major posttranslational modification of proteins. The level of O-glycosylation at a site could be useful in terms of evaluating various disease conditions. To address the feasibility of measuring O-glycosylation levels based on the glycopeptide ion intensity in a mass spectrum, apoliporotein CIII (apoC3), a protein that contains a single core-1 O-glycan Gal–GalNAc disaccharide was analyzed by matrix-assisted laser desorption ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS). The intensity of protonated ions for an equimolar mixture of desialylated and deglycosylated apoC3s were the same in linear TOF measurements. No substantial in-source decay, including the cleavage of the protein-sugar linkage was observed. The glycopeptide derived from apoC3 and the unglycosylated counterpart, when analyzed by MALDI reflectron TOF MS indicated that post-source decay was minimal. These collective findings demonstrate the feasibility of label-free quantitation of O-glycan occupancy by MS when the glycans are small and neutral. This method provides a tool for use in glycoproteomics as a complement of our previous report (DOI: 10.1021/pr900913k) for calculating the saccharide composition of O-glycans.

収録刊行物

• Mass Spectrometry

  Mass Spectrometry 1 (2), A0008-A0008, 2012

  一般社団法人　日本質量分析学会

キーワード

• O-glycan
• apolipoprotein CIII
• matrix-assisted laser desorption/ionization
• label-free quantitation
• in-source decay

詳細情報

• CRID: 1390282680392180736
• NII論文ID: 10031126345
• NII書誌ID: AN0010555X
• ISSN: 13408097
• ISSN: 21865116
• ISSN: 2187137X
• DOI: 10.5702/massspectrometry.a0008
• NDL書誌ID: 024140155
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I024140155
• Web Site: https://www.jstage.jst.go.jp/article/massspectrometry/1/2/1_A0008/_pdf
• 本文言語コード: en
• データソース種別:
  • JaLC
  • NDL
  • Crossref
  • CiNii Articles
  • KAKEN
• 抄録ライセンスフラグ: 使用不可