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The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is brain specific and not due to mannose-6-phosphorylation

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Abstract

Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C which is predominantly expressed in brain myelin and kidney. Due to shedding of the GPI-anchor eNPP6 occurs also as a soluble isoform (s-eNPP6). eNPP 6 consists of two identical monomers of 55 kDa joined by a disulfide bridge, and possesses four N-glycans in each monomer. In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. Here we completed characterization of the site-specific glycan structures of bovine brain s-eNPP6, and determined the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney. Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. Thus, the non-classical glycan processing pathway of brain eNPP 6 is not due to mannose-6-phosphorylation, suggesting that there is an alternative Golgi glycan-processing pathway of eNPP6 in brain. The resulting brain-specific expression of accessible hybrid and oligomannosidic glycans may be physiologically important within the cell–cell communication system of the brain.

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Abbreviations

eNPP:

Ectonucleotide pyrophosphatase/phosphodiesterase

endo H:

Mannosyl-glycoprotein endo-β-N-acetylglucosaminidase (EC 3.2.1.96)

PNGase F:

Peptide-N4-(N-acetyl-β-glucosaminyl)asparagine amidase (EC 3.5.1.52)

GPI:

Glycosylphosphatidylinositol

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Acknowledgments

We thank Wenche Bakkelund for technical support and Toril Anne Gronseth, Department of Pharmacy, University of Tromso for help on Mass Spectrometry analyses. The work was supported by Institute of Medical Biology, the Health Faculty, University of Tromsø.

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  1. Institute of Medical Biology, University of Tromso, MH Building, 9037, Tromso, Norway

    Ole K. Greiner-Tollersrud

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  1. Ole K. Greiner-Tollersrud
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Correspondence to Ole K. Greiner-Tollersrud.

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Greiner-Tollersrud, O.K. The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is brain specific and not due to mannose-6-phosphorylation. Neurochem Res 39, 2025–2029 (2014). https://doi.org/10.1007/s11064-014-1412-1

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