Modification of sialic acid metabolism of murine erythroleukemia cells by analogs of N-acetylmannosamine

Edward L. Schwartz, Anthony F. Hadfield, Alfred E. Brown, Alan C. Sartorelli

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Two analogs of N-acetylmannosamine, 2-acetamido-1,3,4,6-tetra-O-acetyl-2-deoxy-d-mannopyranose (Ac4-NAcMan) and the 2-trifluoroacetamido derivative (Ac4F3-NAcMan), were synthesized as potential inhibitors of the formation of sialic acid-containing glycoconjugates and were examined for their ability to modify the incorporation of N-[3H]acetylmannosamine into cellular glycoconjugates of Friend murine erythroleukemia cells. Ac4F3-NAcMan and Ac4-NAcMan inhibited cellular replication in suspension culture at concentrations of 0.02 and 0.08 mM, respectively. The cytotoxicity of Ac4-NAcMan was relatively reversible, whereas that produced by Ac4F3-NAcMan was not, as judged by measurement of the cloning efficiencies of cells exposed to these agents. The analogs inhibited incorporation of N-[3H]acetylmannosamine into ethanol-soluble and -insoluble materials. Separation of ethanol-soluble metabolites by HPLC demonstrated that Ac4F3-NAcMan caused accumulation of radioactivity from N-[3H]acetylmannosamine in CMP-N-acetylneuraminic acid (CMP-NeuNAc) equal to the decrease in macromolecular-bound 3H caused by this agent. In contrast, similar exposure to Ac4-NAcMan produced a large increase in the amount of radioactivity in ethanol-soluble N-acetylneuraminic acid while decreasing the amount of label from N-[3H]acetylmannosamine in cellular CMP-NeuNAc, suggesting that the analogs differ in their biochemical sites of action. Treatment of cells with either analog increased the amount of neuraminidase-hydrolyzable sialic acid-like material on the cell surface; this appeared to be due to the incorporation of the analogs into cellular glycoconjugates, since incubation of cells with 3H-labeled analogs resulted in the appearance of radioactivity in cellular ethanol-insoluble and neuraminidase-hydrolyzable material. Incubation of cells with Ac4-NAcMan labeled with 14C in the 4-O-acetyl group further demonstrated that incorporation occurred with approx. 50% retention of this substituent. Thus, both the amount and the nature of the surface sialic acid constituents of treated cells were altered, suggesting that these or similar analogs could potentially be used to modify cellular membrane function.

Original languageEnglish (US)
Pages (from-to)489-497
Number of pages9
JournalBBA - Molecular Cell Research
Volume762
Issue number4
DOIs
StatePublished - Jul 14 1983
Externally publishedYes

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Leukemia, Erythroblastic, Acute
N-Acetylneuraminic Acid
Cytidine Monophosphate
Glycoconjugates
Ethanol
Radioactivity
Neuraminidase
Mannose
N-acetylmannosamine
Organism Cloning
Suspensions
High Pressure Liquid Chromatography
2-acetamido-1,3,4,6-tetra-O-acetyl-2-deoxymannopyranose
Membranes
2-trifluoroacetamido-1,3,4,6-tetra-O-acetyl-2-deoxymannopyranose

Keywords

  • (Friend cell)
  • Acetylmannosamine analog
  • Sialic acid metabolism

ASJC Scopus subject areas

  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Modification of sialic acid metabolism of murine erythroleukemia cells by analogs of N-acetylmannosamine. / Schwartz, Edward L.; Hadfield, Anthony F.; Brown, Alfred E.; Sartorelli, Alan C.

In: BBA - Molecular Cell Research, Vol. 762, No. 4, 14.07.1983, p. 489-497.

Research output: Contribution to journalArticle

Schwartz, Edward L. ; Hadfield, Anthony F. ; Brown, Alfred E. ; Sartorelli, Alan C. / Modification of sialic acid metabolism of murine erythroleukemia cells by analogs of N-acetylmannosamine. In: BBA - Molecular Cell Research. 1983 ; Vol. 762, No. 4. pp. 489-497.
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N2 - Two analogs of N-acetylmannosamine, 2-acetamido-1,3,4,6-tetra-O-acetyl-2-deoxy-d-mannopyranose (Ac4-NAcMan) and the 2-trifluoroacetamido derivative (Ac4F3-NAcMan), were synthesized as potential inhibitors of the formation of sialic acid-containing glycoconjugates and were examined for their ability to modify the incorporation of N-[3H]acetylmannosamine into cellular glycoconjugates of Friend murine erythroleukemia cells. Ac4F3-NAcMan and Ac4-NAcMan inhibited cellular replication in suspension culture at concentrations of 0.02 and 0.08 mM, respectively. The cytotoxicity of Ac4-NAcMan was relatively reversible, whereas that produced by Ac4F3-NAcMan was not, as judged by measurement of the cloning efficiencies of cells exposed to these agents. The analogs inhibited incorporation of N-[3H]acetylmannosamine into ethanol-soluble and -insoluble materials. Separation of ethanol-soluble metabolites by HPLC demonstrated that Ac4F3-NAcMan caused accumulation of radioactivity from N-[3H]acetylmannosamine in CMP-N-acetylneuraminic acid (CMP-NeuNAc) equal to the decrease in macromolecular-bound 3H caused by this agent. In contrast, similar exposure to Ac4-NAcMan produced a large increase in the amount of radioactivity in ethanol-soluble N-acetylneuraminic acid while decreasing the amount of label from N-[3H]acetylmannosamine in cellular CMP-NeuNAc, suggesting that the analogs differ in their biochemical sites of action. Treatment of cells with either analog increased the amount of neuraminidase-hydrolyzable sialic acid-like material on the cell surface; this appeared to be due to the incorporation of the analogs into cellular glycoconjugates, since incubation of cells with 3H-labeled analogs resulted in the appearance of radioactivity in cellular ethanol-insoluble and neuraminidase-hydrolyzable material. Incubation of cells with Ac4-NAcMan labeled with 14C in the 4-O-acetyl group further demonstrated that incorporation occurred with approx. 50% retention of this substituent. Thus, both the amount and the nature of the surface sialic acid constituents of treated cells were altered, suggesting that these or similar analogs could potentially be used to modify cellular membrane function.

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