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Glycoconjugate Journal

, Volume 10, Issue 1, pp 109–115 | Cite as

Regulation of biosynthesis ofN-glycolylneuraminic acid-containing glycoconjugates: characterization of factors required for NADH-dependent cytidine 5′monophosphate-N-acetylneuraminic acid hydroxylation

  • Takehiro Kawano
  • Yasunori Kozutsumi
  • Hiromu Takematsu
  • Toshisuke Kawasaki
  • Akemi Suzuki
Papers

Abstract

The hydroxylation of CMP-NeuAc has been demonstrated to be carried out by several factors including the soluble form of cytochromeb5. In the present study, mouse liver cytosol was subjected to ammonium sulfate fractionation and cellulose phosphate column chromatography for the separation of two other essential fractions participating in the hydroxylation. One of the fractions, which bound to a cellulose phosphate column, was able to reduce the soluble cytochromeb5, using NADH as an electron donor. The other fraction, which flowed through the column, was assumed to contain the terminal enzyme which accepts electrons from cytochromeb5, activates oxygen, and catalyses the hydroxylation of CMP-NeuAc. Assay conditions for the quantitative determination of the terminal enzyme were established, and the activity of the enzyme in several tissues of mouse and rat was measured. The level of the terminal enzyme activity is associated with the expression ofN-glycolylneuraminic acid in these tissues, indicating that the expression of the terminal enzyme possibly regulates the overall velocity of CMP-NeuAc hydroxylation.

Keywords

CMP-NeuAc hydroxylase N-glycolylneuraminic acid H-D antigen cytochromeb5 

Abbreviations

CMP

cytidine 5′-monophosphate

NeuAc

N-acetylneuraminic acid

NeuGc

N-glycolylneuraminic acid

NADH

reduced nicotinamide adenine dinucleotide

NADPH

reduced nicotinamide adenine dinucleotide phosphate

DTT

dithiothreitol

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Copyright information

© Chapman & Hall 1993

Authors and Affiliations

  • Takehiro Kawano
    • 1
  • Yasunori Kozutsumi
    • 2
  • Hiromu Takematsu
    • 2
  • Toshisuke Kawasaki
    • 2
  • Akemi Suzuki
    • 1
  1. 1.Department of Membrane BiochemistryTokyo Metropolitan Institute of Medical ScienceTokyoJapan
  2. 2.Department of Biological Chemistry, Faculty of Pharmaceutical SciencesKyoto UniversityKyotoJapan

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