Open AccessDetermination of the structure of the carbohydrate chains of prostaglandin endoperoxide synthase from sheep
Author(s) -
MUTSAERS Johanna H. G. M.,
HALBEEK Herman,
KAMERLING Johannis P.,
VLIEGENTHART Johannes F. G.
Publication year - 1985
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.0014-2956.1985.00569.x
Subject(s) - atp synthase , chemistry , carbohydrate , biochemistry , prostaglandin , stereochemistry , enzyme
Prostaglandin endoperoxide synthase was isolated from sheep seminal vesicles. Sugar analysis of the glycoprotein revealed the presence of mannose and N ‐acetylglucosamine only. The carbohydrate moiety was released from the polypeptide backbone by hydrazinolysis. After re‐ N ‐acetylation and reduction, the resulting mixture of oligosaccharide‐alditols was fractionated on Bio‐Gel P‐4 and their structures were investigated by 500‐MHz 1 H‐NMR spectroscopy. The carbohydrate chains turned out to be of the oligomannoside type containing six to nine mannose residues. The largest and most abundant compound was established to be:For the smaller structures heterogeneity occurs with respect to the outer α(1 → 2)‐linked mannose residues. Furthermore, a small amount of Man 6 GlcNAc‐ol (artefact of the hydrazinolysis procedure) was detected by 1 H‐NMR spectroscopy and fast atom bombardment mass spectrometry.