Characterization of a new type of glycoprotein saccharides containing polysialosyl sequence

doi:10.1016/S0006-291X(80)80260-9

Biochemical and Biophysical Research Communications

Volume 93, Issue 1, 13 March 1980, Pages 162-165

https://doi.org/10.1016/S0006-291X(80)80260-9 Get rights and content

Summary

The occurrence of polysialosyl sequence has been shown for the first time in the carbohydrate units of glycoprotein isolated from the eggs of rainbow trout. The saccharide chains accounting for 80 % of the weight of the glycoprotein were O-glycosidically linked to the peptide moiety. The reduced saccharides obtained by alkaline borohydride treatment of the glycoprotein contain various amounts of N-glycolylneuraminic acid and were fractionated by a column of DEAE-Sephadex. Analyses of the saccharides containing large amounts of N-glycolylneuraminic acid show that 2 → 8 linked homopolymers of more than 15 N-glycolylneuraminic acid residues were attached to the core composed of 2 galactose, 1 N-acetylgalactosamine, and 1 N-acetylgalactosaminitol residues.

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Cited by (48)

Metabolism of vertebrate amino sugars with N-glycolyl groups: Resistance of α2-8-linked N-glycolylneuraminic acid to enzymatic cleavage
2012, Journal of Biological Chemistry
Citation Excerpt :
However, no bacterium has yet been shown to be capable of Neu5Gc synthesis, and thus these capsules contain only Neu5Ac. There is a single report of Neu5Gc present in α2–8-linked polySia is in the eggs of salmonid fish (113). Although we are not aware of any organism that digests these glycoproteins, we can hypothesize that eggs that contain Neu5Gc may be more resistant to attack by sialidase-producing pathogens in the wild.
The sialic acid (Sia) N-acetylneuraminic acid (Neu5Ac) and its hydroxylated derivative N-glycolylneuraminic acid (Neu5Gc) differ by one oxygen atom. CMP-Neu5Gc is synthesized from CMP-Neu5Ac, with Neu5Gc representing a highly variable fraction of total Sias in various tissues and among different species. The exception may be the brain, where Neu5Ac is abundant and Neu5Gc is reported to be rare. Here, we confirm this unusual pattern and its evolutionary conservation in additional samples from various species, concluding that brain Neu5Gc expression has been maintained at extremely low levels over hundreds of millions of years of vertebrate evolution. Most explanations for this pattern do not require maintaining neural Neu5Gc at such low levels. We hypothesized that resistance of α2–8-linked Neu5Gc to vertebrate sialidases is the detrimental effect requiring the relative absence of Neu5Gc from brain. This linkage is prominent in polysialic acid (polySia), a molecule with critical roles in vertebrate neural development. We show that Neu5Gc is incorporated into neural polySia and does not cause in vitro toxicity. Synthetic polymers of Neu5Ac and Neu5Gc showed that mammalian and bacterial sialidases are much less able to hydrolyze α2–8-linked Neu5Gc at the nonreducing terminus. Notably, this difference was not seen with acid-catalyzed hydrolysis of polySias. Molecular dynamics modeling indicates that differences in the three-dimensional conformation of terminal saccharides may partly explain reduced enzymatic activity. In keeping with this, polymers of N-propionylneuraminic acid are sensitive to sialidases. Resistance of Neu5Gc-containing polySia to sialidases provides a potential explanation for the rarity of Neu5Gc in the vertebrate brain.
Discovery of an α2,9-PolyNeu5Ac glycoprotein in C-1300 murine neuroblastoma (clone NB41A3)
2003, Journal of Biological Chemistry
α2,8-PolyNeu5Ac is expressed on neural cell adhesion molecules during embryogenesis and also re-expressed on certain tumors. PolyNeu5Ac is therefore an oncodevelopmental antigen, has important regulatory effects on the adhesive and migratory behavior of neural cells, and is thus crucial to synaptic plasticity. Until now, α2,9-polyNeu5Ac, a linkage isomer of α2,8-polyNeu5Ac, has long been thought to occur only in capsules of neuroinvasive Neisseria meningitidis group C bacteria. Here we report the unexpected discovery of α2,9-polyNeu5Ac in a new cell adhesion-related glycoprotein on the membrane of C-1300 murine neuroblastoma cells (clone NB41A3). We also report the expression of α2,9-polyNeu5Ac was affected by cell growth and retinoic acid-induced differentiation. Occurrence of the linkage isomer of α2,8-polyNeu5Ac has been left unrecognized by conventional methods using biological diagnostic probes for α2,8-polyNeu5Ac. Thus, our discovery may change contemporary views of biology and pathology of polysialic acid and open new avenues for the development of anti-neural tumor drugs.
A challenge to the ultrasensitive chemical method for the analysis of oligo- and polysialic acids at a nanogram level of colominic acid and a milligram level of brain tissues
2001, Biochimie
Polysialic acid (polySia) is a functional epitope and is known: 1) to regulate normal fertilization of lower vertebrates and invertebrates; 2) to be expressed on neural cell adhesion molecule (NCAM) when the formation or re-arrangement of nervous tissues takes place during embryonic stages as well as in adults of higher vertebrates; and 3) to be re-expressed in several human tumors. Thus, polySia serves as oncodevelopmental antigen. To date sensitive biochemical diagnostic probes (antibodies and endo-N-acylneuraminidase) to detect polySia are known. However, these reagents are not commercially available yet and they are only reactive to specific types of polySia structure. Moreover, precise information not only on diversity but also on the length or degree of polymerization (DP) of extended polySia chains is considered important in understanding the molecular mechanism of biosynthesis of polySia chains and fine-tuning of NCAM-NCAM adhesive interaction by polySia chain but cannot be obtained with these biochemical probes. We have been continuously making efforts to develop and improve the sensitivity of chemical methods for polySia analysis toward these challenging problems. This article presents our most recently developed chemical method for polySia analysis and its use in obtaining new information on DP of colominic acid samples and polySia chains present in rat brain tissues with the highest sensitivity that has ever been attained.
High-performance capillary electrophoretic characterization of different types of oligo- and polysialic acid chains
1998, Analytical Biochemistry
We have carried out comparative structural analysis of novel oligo- and polysialic acid chains from diverse sources. Controlled acid hydrolysates of (a) colominic acid, α2→8-linked homopolymer ofN-acetylneuraminic acid (Neu5Ac), (b) α2→8-linked oligo/polyNeu5Gc chains present in rainbow trout egg polysialoglycoprotein, and (c) α2→8-linked oligomers of deaminoneuraminic acid (KDN) residues of KDN-rich glycoprotein derived from rainbow trout vitelline envelope were analyzed by high-performance capillary electrophoresis (HPCE). The results showed that three different types of α2→8-linked oligosialic acids having same degree of polymerization can be separated by HPCE. A partial hydrolysate of colominic acid with mild acid was shown by CE to form intramolecular esters during the controlled hydrolysis and the subsequent workup procedure. In contrast, lactonization of (→5-O_glycolyl-Neu5Gcα2→)n, α2→5-O_glycolyl-linked homopolymer ofN-glycolylneuraminic acid (Neu5Gc) present in the egg jelly coat of sea urchin, did not take place as readily as in (→8Neu5Acα2→)n.
An expeditious route to N-glycolylneuraminic acid based on enzyme-catalyzed reaction
1997, Tetrahedron
A new preparative way of N-glycolylneuraminic acid (NeuGc), one of the important family of sialic acids, from N-acetylglucosamine (GlcNAc) via N-acetylmannosamine (ManNAc) was established based on the combination of chemical and enzymatic reactions. In a kinetic study of the key enzymatic reaction for this process, aldolase-catalyzed synthesis of sialic acid, an inhibitory effect of gluco-isomer on the enzymatic reaction was quantitatively clarified, and the importance of isomerically pure substrate with manno-configuration for aldolase-catalyzed reaction was suggested. A newly developed method, selective degradation of GlcNAc contaminating in the substrate by use of Rhodococcus rhodochrous IFO 15564 provided pure ManNAc to avoid such inhibitory effect of the gluco-isomer for aldolase. Starting from pure ManNAc, via mannosamine hydrochloride, acetoxyacetyl chloride was applied for introducing a protected form of glycolyl group to give N-acetylglycolylmannosamine. For the removal of acetyl protective group, a lipase from Aspergillus niger was effectively used under a mild and neutral condition to afford N-glycolylmannosamine (ManNGc), the substrate of aldolase. NeuGc was prepared in 25% yield and 7 steps from GlcNAc.
Chapter 7 Fish Glycoproteins
1997, New Comprehensive Biochemistry

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Characterization of a new type of glycoprotein saccharides containing polysialosyl sequence

Summary

Biochem. Biophys. Res. Commun.

Carbohydr. Res.

J. Biol. Chem.