Luminescence detection of peptide:N-glycanase activity using engineered split inteins

Tsuyoshi Takahashi; Tatsuya Uchibayashi; Nozomi Ishii; Ichiro Matsuo; Yukiko Yoshida; Tadashi Suzuki

doi:10.1039/D2CC04865E

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Luminescence detection of peptide:N-glycanase activity using engineered split inteins†

Tsuyoshi Takahashi,

*^a Tatsuya Uchibayashi,^a Nozomi Ishii,^a Ichiro Matsuo,

^a Yukiko Yoshida

^b and Tadashi Suzuki^c

Author affiliations

* Corresponding authors

^a Graduate School of Science and Technology, Gunma University, 1-5-1 Tenjin-cho, Kiryu, Gunma 376-8515, Japan
E-mail: ttakahas@gunma-u.ac.jp

^b Ubiquitin Project, Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo 156-8506, Japan

^c Glycometabolic Biochemistry Laboratory, RIKEN Cluster for Pioneering Research, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan

Abstract

A split intein-based method has been developed to detect peptide:N-glycanase (PNGase) activity in live cells. PNGase cleaves the linkage between N,N′-diacetylchitobiose and the Asn side-chain of N-intein peptides and the products react rapidly with C-intein by protein trans-splicing to generate an active luciferase.

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Article information

DOI: https://doi.org/10.1039/D2CC04865E
Article type: Communication
Submitted: 02 Sep 2022
Accepted: 07 Nov 2022
First published: 08 Nov 2022

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Chem. Commun., 2022,58, 13282-13285

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Luminescence detection of peptide:N-glycanase activity using engineered split inteins

T. Takahashi, T. Uchibayashi, N. Ishii, I. Matsuo, Y. Yoshida and T. Suzuki, Chem. Commun., 2022, 58, 13282 DOI: 10.1039/D2CC04865E

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Chemical Communications

Luminescence detection of peptide:N-glycanase activity using engineered split inteins†

Abstract

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Luminescence detection of peptide:N-glycanase activity using engineered split inteins

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