Improved assay system for acidic peptide: N-glycanase (aPNGase) activity in plant extracts

Chiharu Yamamoto, Mikako Ogura, Ryota Uemura, Maeda Megumi, Hiroyuki Kajiura, Ryo Misaki, Kazuhito Fujiyama, Yoshinobu Kimura

Research output: Contribution to journalArticlepeer-review

Abstract

Plant acidic peptide: N-glycanase (aPNGase) release N-glycans from glycopeptides during the degradation process of glycoproteins in developing or growing plants. We have previously developed a new method to detect the aPNGase activity in crude extracts, which is prerequisite for the construction of aPNGase knockout or overexpression lines. However, this method has the disadvantage of requiring de-sialylation treatment and a lectin chromatography. In this study, therefore, we improved the simple and accurate method for detecting aPNGase activity using anion-exchange HPLC requiring neither the desialylation treatment nor the lectin affinity chromatography.

Original languageEnglish
Article number114367
JournalAnalytical Biochemistry
Volume634
DOIs
Publication statusPublished - Dec 1 2021

Keywords

  • Acidic PNGase
  • Arabidopsis thaliana
  • Free N-Glycans
  • PNGase assay
  • Transgenic plant

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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