Document Detail


Use of prokaryotic-derived probes to identify poly(sialic acid) in neonatal neuronal membranes.
MedLine Citation:
PMID:  6371806     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Three prokaryotic-derived probes to identify and study the temporal expression of polysialosyl units in neuronal tissue have been developed. A polyclonal antibody, a bacteriophage-derived endo-neuraminidase, and an Escherichia coli K1 sialyltransferase are all specific for either recognizing or synthesizing poly(sialic acid) containing alpha-2,8-ketosidic linkages. Polysialosyl immunoreactivity with apparent Mr values of 180,000-240,000 was specific for developing neuronal tissue; it was not detected in neonatal liver or kidney or in adult brain tissue. The developmentally regulated disappearance in poly(sialic acid) is consistent with the probes described here recognizing the polysialosyl carbohydrate units of a neuronal cell adhesion molecule (N-CAM). Treatment of brain extracts with a bacteriophage-derived endo-neuraminidase specific for alpha-2,8-linked polysialosyl units abolished the immunoreactivity. The material solubilized by endo-neuraminidase was isolated, reduced with borotritide, and shown to contain oligomers of sialic acid with three to six sialyl units. Treatment of the 3H-labeled oligosialic acid with exo-neuraminidase quantitatively converted the radioactivity to sialitol, establishing that the brain-derived oligomers were composed solely of sialic acid. A membranous sialytransferase from E. coli K1 that can transfer sialic acid to exogenous acceptors of oligo- or poly(sialic acid) also recognized rat brain membranes, further substantiating the presence of poly(sialic acid) in rat brain. This conclusion was confirmed by using a mutant of E. coli K1 that was defective in the synthesis of poly(sialic acid) and could only transfer sialic acid to exogenous acceptors of oligo- or poly(sialic acid). Sialyl polymer synthesis was restored in the mutant when brain membranes were added as exogenous acceptor.
Authors:
E R Vimr; R D McCoy; H F Vollger; N C Wilkison; F A Troy
Related Documents :
628316 - Composition and synthesis of three higher ganglioside homologs in bovine mammary tissue.
16847056 - Isoform-specific effects of the beta2 subunit on voltage-gated sodium channel gating.
7849276 - Carbohydrates of pigeon milk and their changes in the first week of secretion.
14736726 - Detailed structural features of glycan chains derived from alpha1-acid glycoproteins of...
9335296 - Structural characterization of the lipid a component of helicobacter pylori rough- and ...
241696 - The ph-dependence of the peptidase activity of aminoacylase.
Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  81     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  1984 Apr 
Date Detail:
Created Date:  1984-05-30     Completed Date:  1984-05-30     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1971-5     Citation Subset:  IM    
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Animals
Brain Chemistry*
Cell Membrane / analysis
Escherichia coli / enzymology
Kinetics
Neuraminidase
Neurons / analysis*
Polysaccharides / analysis*
Rats
Sialic Acids / analysis*
Sialyltransferases / metabolism
Grant Support
ID/Acronym/Agency:
AI-09352/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Polysaccharides; 0/Sialic Acids; 0/polysialic acid; EC 2.4.99.-/Sialyltransferases; EC 3.2.1.18/Neuraminidase
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Strategy for the mass spectrometric verification and correction of the primary structures of protein...
Next Document:  Structure of human hemopexin: O-glycosyl and N-glycosyl sites and unusual clustering of tryptophan r...