Structural characterization of glycopeptides by N-terminal protein ladder sequencing |
| |
Authors: | Suzuki Yusuke Suzuki Minoru Nakahara Yoshiaki Ito Yukishige Ito Emi Goto Naoko Miseki Kozo Iida Junko Suzuki Akemi |
| |
Affiliation: | Sphingolipid Expression Laboratory, Supra Biomolecular System Research Group, RIKEN Frontier Research System, Wako-shi, Saitama 351-0198, Japan. |
| |
Abstract: | High-sensitivity and high-throughput mass spectrometry (MS) has become an important tool for characterizing glycopeptides. Here, we analyzed synthetic O-linked glycopeptides using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS. First, we applied MALDI-quadrupole ion trap (QIT)-TOF MS, which enables collision-induced dissociation-MSn analysis for fine structural characterization. Subsequent MS/MS of sodium adduct ions selected as precursor ions yielded detailed information about the site of oligosaccharide attachment as well as the carbohydrate and amino acid sequences; however, these MS/MS spectra were very complex. To obtain easily interpretable and simple spectra, we used N-terminal protein ladder sequencing coupled with MALDI-TOF MS. From the extremely simple resulting spectra, we were able to determine the glycosylation sites, amino acid sequences, and oligosaccharide molecular weights of the glycopeptides. |
| |
Keywords: | |
本文献已被 PubMed 等数据库收录! |
|