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Poster Presentations
Day 2, June 23（Mon.）
Room P (Maesato East, Foyer, Ocean Wing)
- 2P-PM-18
MALDI-TOF MS analysis of unique O- and N-glycans on proteins in unfertilized salmon eggs using aminolysis-SALSA method
(1Nagoya Univ. 1, 2Nagoya Univ. 2)
oMasaki Kurogochi1, Kai Suzuki1,2, Di Wu1,2, Hisatoshi Hanamatsu1, Ken Kitajima1, Chihiro Sato1,2, Jun-ichi Furukawa1
Previously, we developed the method of sialic acid linkage-specific alkylamidation via lactone ring-opening aminolysis (aminolysis-SALSA) for the distinguishment of the linkage of sialic acid; however, it is not clear whether oligo- and poly-sialic acids are derivatized or not by aminolysis-SALSA. It is known that O-glycans containing oligosialic acids with α2,8 linkages are present in unfertilized and fertilized eggs of salmon and rainbow trout. In this study, we investigated the analysis of O- and N-glycans in unfertilized and fertilized eggs of salmon using an aminolysis-SALSA method. First, the carboxyl groups of sialic acids of O-glycans on proteins were directly amidated by in-solution SALSA. Next, SALSA-derivatized O-glycans were cleaved and labeled with 1-phenyl-3-methyl-5-pyrazolone using an evaporative α-elimination, and analyzed by MALDI-TOF MS. On the other hand, N-glycans were cleaved with PNGase F, captured on hydrazide beads by chemical ligation (glycoblotting method), derivatized by SALSA on solid-phase, released and labeled with benzyloxyamine (BOA), and then analyzed by MALDI-TOF MS.