Open AccessMolecular characterization of Limulus Polyphemus C‐reactive protein
Author(s) -
AMATAYAKULCHANTLER Supavadee,
DWEK Raymond A.,
TENNENT Glenys A.,
PEPYS Mark B.,
RADEMACHER Thomas W.
Publication year - 1993
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1993.tb17901.x
Subject(s) - limulus , polyphemus , horseshoe crab , characterization (materials science) , chemistry , computational biology , biology , materials science , evolutionary biology , nanotechnology , ecology
The N‐linked oligosaccharides of C‐reactive protein (CRP) from the arachnid Limulus polyphemus , the horsehoe crab, were characterized after their release by hydrazinolysis, re‐ N ‐acetylation, and reduction with NaB 3 H 4 . High‐voltage paper electrophoresis of the reduced oligosaccharides revealed only neutral species. Gel‐permeation chromatography on Bio‐Gel P4 yielded five fractions. The oligosaccharide fractions were further fractionated using high‐voltage borate paper electrophoresis and Dionex BioLC ion‐exchange chromatography. The oligosaccharides were structurally characterized by sequential exoglycosidase digestion, fragmentation by acetolysis and methylation analysis. Three major structures were found, of which two were the biantennary oligomannose type with compositions Man 5 GlcNAc 2 (B‐1), Man 4 GlcNAc 2 (C‐3) and one was the monoantennary structure Man 3 GlcNAc 2 ( d ‐1). The biantennary oligomannose structures B‐1 and C‐3 contained the structural unit Manα6Manα6R. This unusual arrangement of mannose linkages suggests a biosynthetic pathway in Limulus which differs from that reported in mammals, plants and the parasitic protozoa.