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Differential Labeling of Glycoproteins with Alkynyl Fucose Analogs
Fucosylated glycans critically regulate the physiological functions of proteins and cells. Alterations in levels of fucosylated glycans are associated with various diseases. For detection and functional modulation of fucosylated glycans, chemical biology approaches using fucose (Fuc) analogs are use...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7503990/ https://www.ncbi.nlm.nih.gov/pubmed/32825463 http://dx.doi.org/10.3390/ijms21176007 |
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author | Ma, Chenyu Takeuchi, Hideyuki Hao, Huilin Yonekawa, Chizuko Nakajima, Kazuki Nagae, Masamichi Okajima, Tetsuya Haltiwanger, Robert S. Kizuka, Yasuhiko |
author_facet | Ma, Chenyu Takeuchi, Hideyuki Hao, Huilin Yonekawa, Chizuko Nakajima, Kazuki Nagae, Masamichi Okajima, Tetsuya Haltiwanger, Robert S. Kizuka, Yasuhiko |
author_sort | Ma, Chenyu |
collection | PubMed |
description | Fucosylated glycans critically regulate the physiological functions of proteins and cells. Alterations in levels of fucosylated glycans are associated with various diseases. For detection and functional modulation of fucosylated glycans, chemical biology approaches using fucose (Fuc) analogs are useful. However, little is known about how efficiently each unnatural Fuc analog is utilized by enzymes in the biosynthetic pathway of fucosylated glycans. We show here that three clickable Fuc analogs with similar but distinct structures labeled cellular glycans with different efficiency and protein specificity. For instance, 6-alkynyl (Alk)-Fuc modified O-Fuc glycans much more efficiently than 7-Alk-Fuc. The level of GDP-6-Alk-Fuc produced in cells was also higher than that of GDP-7-Alk-Fuc. Comprehensive in vitro fucosyltransferase assays revealed that 7-Alk-Fuc is commonly tolerated by most fucosyltransferases. Surprisingly, both protein O-fucosyltransferases (POFUTs) could transfer all Fuc analogs in vitro, likely because POFUT structures have a larger space around their Fuc binding sites. These findings demonstrate that labeling and detection of fucosylated glycans with Fuc analogs depend on multiple cellular steps, including conversion to GDP form, transport into the ER or Golgi, and utilization by each fucosyltransferase, providing insights into design of novel sugar analogs for specific detection of target glycans or inhibition of their functions. |
format | Online Article Text |
id | pubmed-7503990 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-75039902020-09-24 Differential Labeling of Glycoproteins with Alkynyl Fucose Analogs Ma, Chenyu Takeuchi, Hideyuki Hao, Huilin Yonekawa, Chizuko Nakajima, Kazuki Nagae, Masamichi Okajima, Tetsuya Haltiwanger, Robert S. Kizuka, Yasuhiko Int J Mol Sci Article Fucosylated glycans critically regulate the physiological functions of proteins and cells. Alterations in levels of fucosylated glycans are associated with various diseases. For detection and functional modulation of fucosylated glycans, chemical biology approaches using fucose (Fuc) analogs are useful. However, little is known about how efficiently each unnatural Fuc analog is utilized by enzymes in the biosynthetic pathway of fucosylated glycans. We show here that three clickable Fuc analogs with similar but distinct structures labeled cellular glycans with different efficiency and protein specificity. For instance, 6-alkynyl (Alk)-Fuc modified O-Fuc glycans much more efficiently than 7-Alk-Fuc. The level of GDP-6-Alk-Fuc produced in cells was also higher than that of GDP-7-Alk-Fuc. Comprehensive in vitro fucosyltransferase assays revealed that 7-Alk-Fuc is commonly tolerated by most fucosyltransferases. Surprisingly, both protein O-fucosyltransferases (POFUTs) could transfer all Fuc analogs in vitro, likely because POFUT structures have a larger space around their Fuc binding sites. These findings demonstrate that labeling and detection of fucosylated glycans with Fuc analogs depend on multiple cellular steps, including conversion to GDP form, transport into the ER or Golgi, and utilization by each fucosyltransferase, providing insights into design of novel sugar analogs for specific detection of target glycans or inhibition of their functions. MDPI 2020-08-20 /pmc/articles/PMC7503990/ /pubmed/32825463 http://dx.doi.org/10.3390/ijms21176007 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Ma, Chenyu Takeuchi, Hideyuki Hao, Huilin Yonekawa, Chizuko Nakajima, Kazuki Nagae, Masamichi Okajima, Tetsuya Haltiwanger, Robert S. Kizuka, Yasuhiko Differential Labeling of Glycoproteins with Alkynyl Fucose Analogs |
title | Differential Labeling of Glycoproteins with Alkynyl Fucose Analogs |
title_full | Differential Labeling of Glycoproteins with Alkynyl Fucose Analogs |
title_fullStr | Differential Labeling of Glycoproteins with Alkynyl Fucose Analogs |
title_full_unstemmed | Differential Labeling of Glycoproteins with Alkynyl Fucose Analogs |
title_short | Differential Labeling of Glycoproteins with Alkynyl Fucose Analogs |
title_sort | differential labeling of glycoproteins with alkynyl fucose analogs |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7503990/ https://www.ncbi.nlm.nih.gov/pubmed/32825463 http://dx.doi.org/10.3390/ijms21176007 |
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