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Glycoconjugate Journal

, Volume 11, Issue 3, pp 194–203 | Cite as

Purification, characterization and reconstitution of CMP-N-acetylneuraminate hydroxylase from mouse liver

  • Petra Schneckenburger
  • Lee Shaw
  • Roland Schauer
Article

Abstract

CMP-N-acetylneuraminate hydroxylase was isolated from mouse liver high speed supernatant with a yield of 0.4% and an apparent 1000-fold purification. The enzyme is a monomeric protein with a molecular weight of 66 kDa, as determined by gel filtration and SDS-PAGE. The hydroxylase system was reconstituted with Triton X-100-solubilized mouse liver microsomes and purified soluble or microsomal forms of cytochrome b5 reductase and cytochrome b5. The systems were characterized in detail and kinetic parameters for each system were determined.

Keywords

sialic acid N-glycoloylneuraminic acid hydroxylase protein purification cytochrome b5 electron transfer enzyme system reconstitution 

Abbreviations

Neu5Ac

N-acetyl-β-d-neuraminic acid

Neu5Gc

N-glycoloyl-β-d-neuraminic acid

CMP-Neu5Ac

cytidine-5′-monophospho-N-acetylneuraminic acid

CMP-Neu5Gc

cytidine-5′-monophospho-N-glycoloylneuraminic acid

TCA

trichloroacetic acid

Chaps

3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulphonate

SOD

superoxide dismutase

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Copyright information

© Chapman & Hall 1994

Authors and Affiliations

  • Petra Schneckenburger
    • 1
  • Lee Shaw
    • 1
  • Roland Schauer
    • 1
  1. 1.Biochemisches Institut der Christian-Albrechts-Universität zu KielKielFRG

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