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Identification, characterization, and developmental expression of a novel α2 → 8-KDN-transferase which terminates elongation of α2 → 8-linked oligo-polysialic acid chain synthesis in trout egg polysialoglycoproteins

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Abstract

A novel glycosyltransferase which catalyses transfer of deaminated neuraminic acid, KDN (2-keto-3-deoxy-d-glycero-d-galacto-nononic acid) from CMP-KDN to the non-reducing termini of oligo-polysialyl chains of polysialoglycoprotein (PSGP), was discovered in the ovary of rainbow trout (Oncorhynchus mykiss). The KDN-transferase activity was optimal at neutral pH, and stimulated 2 to 2.5-fold by 2–5mm Mg2+ or Mn2+. Expression of KDN-transferase was developmentally regulated in parallel with expression of the α2 → 8-polysialytransferase, which catalyses synthesis of the oligo-polysialyl chains in PSGP. Incorporation of the KDN residues into the oligo-polysialyl chains prevented their further elongation, resulting in ‘capping’ of the oligo-polysialyl chains. This is the first example of a glycosyltransferase that catalyses termination of α2 → 8-polysialylation in glycoproteins.

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Abbreviations

KDN:

2-keto-3-deoxy-d-glycero-d-galacto-nononic acid or naturally occurring deaminated neuraminic acid

Neu5Ac:

N-acetylneuraminic acid

Neu5Ge:

N-glycolylneuraminic acid

CMP-KDN:

cytidine 5′-(3-deoxy-d-glycero-d-galacto-2-nonulosonic phosphate) or cytidine 5′-KDN phosphate

CMP-NeuAc:

cytidine 5′-Neu5Ac phosphate; oligo-polySia, oligo- and/or polysialic acid

PSGP:

rainbow trout egg polysialoglycoprotein comprising α2 → 8-linked oligo- polyNeu5Gc

PSGP (low Sia):

a precursor of PSGP present at early stages of oogenesis which contains mostly the disialyl group, Siaα2 → 8Siaα2 → 6-

*K-PSGP:

[14C]KDN-labelled PSGP obtained by incubating PSGP and CMP-[14C]KDN with the immature cortical vesicle fraction P1 containing KDN-transferase

*A-PSGP:

[14C]Neu5Ac-labelled PSGP obtained by incubating PSGP and CMP-[14C]Neu5Ac with the P1 fraction

A-*K-PSGP andK-*K-PSGP:

the products obtained after incubating *K-PSGP with P1 fraction and unlabelled CMP-Neu5Ac or CMP-KDN, respectively

*K-PSGP cho ,A-*K-PSGP cho , andK-*K-PSGP cho :

mixture of oligosaccharide alditols obtained by alkaline borohydride treatment of *K-PSGP,A-*K-PSGP, and K-*K-PSGP, respectively

*A-PSGP cho :

a mixture of oligosaccharide alditols obtained by alkaline borohydride treatment of [14C]Neu5Ac-labelled PSGP

Endo-N:

endo-N-acylneuraminidase

DP:

degree of polymerization

GLC:

gas-liquid chromatography

HPLC:

high performance liquid chromatography

TLC:

thin layer chromatography

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Authors and Affiliations

  1. Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Hongo-7, 113, Tokyo, Japan

    Takashi Angata, Shinobu Kitazume, Takaho Terada, Ken Kitajima & Yasuo Inoue

  2. School of Pharmaceutical Sciences, Showa University, Hatanodai-1, 142, Tokyo, Japan

    Sadako Inoue

  3. Department of Biological Chemistry, School of Medicine, University of California, 95616, Davis, CA, USA

    Frederic A. Troy II

Authors
  1. Takashi Angata
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  2. Shinobu Kitazume
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  3. Takaho Terada
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  4. Ken Kitajima
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  5. Sadako Inoue
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  6. Frederic A. Troy II
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  7. Yasuo Inoue
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Angata, T., Kitazume, S., Terada, T. et al. Identification, characterization, and developmental expression of a novel α2 → 8-KDN-transferase which terminates elongation of α2 → 8-linked oligo-polysialic acid chain synthesis in trout egg polysialoglycoproteins. Glycoconjugate J 11, 493–499 (1994). https://doi.org/10.1007/BF00731286

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  • DOI: https://doi.org/10.1007/BF00731286

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