Characterization of protein glycosylation in an Asgard archaeon

Archaeal cells are typically enveloped by glycosylated S-layer proteins. Archaeal protein glycosylation provides valuable insights not only into their adaptation to their niches but also into their evolutionary trajectory. Notably, thermophilic Thermoproteota modify proteins with N-glycans that incl...

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Main Authors: Satoshi Nakagawa, Hiroyuki Imachi, Shigeru Shimamura, Saeko Yanaka, Hirokazu Yagi, Maho Yagi-Utsumi, Hiroyuki Sakai, Shingo Kato, Moriya Ohkuma, Koichi Kato, Ken Takai
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Language:English
Published: Elsevier 2024-01-01
Series:BBA Advances
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Online Access:http://www.sciencedirect.com/science/article/pii/S2667160324000061
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author Satoshi Nakagawa
Hiroyuki Imachi
Shigeru Shimamura
Saeko Yanaka
Hirokazu Yagi
Maho Yagi-Utsumi
Hiroyuki Sakai
Shingo Kato
Moriya Ohkuma
Koichi Kato
Ken Takai
author_facet Satoshi Nakagawa
Hiroyuki Imachi
Shigeru Shimamura
Saeko Yanaka
Hirokazu Yagi
Maho Yagi-Utsumi
Hiroyuki Sakai
Shingo Kato
Moriya Ohkuma
Koichi Kato
Ken Takai
author_sort Satoshi Nakagawa
collection DOAJ
description Archaeal cells are typically enveloped by glycosylated S-layer proteins. Archaeal protein glycosylation provides valuable insights not only into their adaptation to their niches but also into their evolutionary trajectory. Notably, thermophilic Thermoproteota modify proteins with N-glycans that include two GlcNAc units at the reducing end, resembling the ''core structure'' preserved across eukaryotes. Recently, Asgard archaea, now classified as members of the phylum Promethearchaeota, have offered unprecedented opportunities for understanding the role of archaea in eukaryogenesis. Despite the presence of genes indicative of protein N-glycosylation in this archaeal group, these have not been experimentally investigated. Here we performed a glycoproteome analysis of the firstly isolated Asgard archaeon Promethearchaeum syntrophicum. Over 700 different proteins were identified through high-resolution LC-MS/MS analysis, however, there was no evidence of either the presence or glycosylation of putative S-layer proteins. Instead, N-glycosylation in this archaeon was primarily observed in an extracellular solute-binding protein, possibly related to chemoreception or transmembrane transport of oligopeptides. The glycan modification occurred on an asparagine residue located within the conserved N-X-S/T sequon, consistent with the pattern found in other archaea, bacteria, and eukaryotes. Unexpectedly, three structurally different N-glycans lacking the conventional core structure were identified in this archaeon, presenting unique compositions that included atypical sugars. Notably, one of these sugars was likely HexNAc modified with a threonine residue, similar to modifications previously observed in mesophilic methanogens within the Methanobacteriati. Our findings advance our understanding of Asgard archaea physiology and evolutionary dynamics.
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spelling doaj-art-9a8e4027d3ef42a599856ca6a754c27e2025-08-20T02:37:53ZengElsevierBBA Advances2667-16032024-01-01610011810.1016/j.bbadva.2024.100118Characterization of protein glycosylation in an Asgard archaeonSatoshi Nakagawa0Hiroyuki Imachi1Shigeru Shimamura2Saeko Yanaka3Hirokazu Yagi4Maho Yagi-Utsumi5Hiroyuki Sakai6Shingo Kato7Moriya Ohkuma8Koichi Kato9Ken Takai10Laboratory of Marine Environmental Microbiology, Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Oiwake-cho, Kitashirakawa, Sakyo-ku, Kyoto 606-8502, Japan; Institute for Extra-cutting-edge Science and Technology Avant-garde Research (X-star), Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima-cho, Yokosuka 273-0061, Japan; Exploratory Research Center on Life and Living Systems (ExCELLS), National Institute of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, Japan; Corresponding author.Institute for Extra-cutting-edge Science and Technology Avant-garde Research (X-star), Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima-cho, Yokosuka 273-0061, JapanInstitute for Extra-cutting-edge Science and Technology Avant-garde Research (X-star), Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima-cho, Yokosuka 273-0061, JapanExploratory Research Center on Life and Living Systems (ExCELLS), National Institute of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, Japan; Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe-dori, Mizuhoku, Nagoya 467-8603, Japan; Institute for Molecular Science (IMS), National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, JapanExploratory Research Center on Life and Living Systems (ExCELLS), National Institute of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, Japan; Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe-dori, Mizuhoku, Nagoya 467-8603, JapanExploratory Research Center on Life and Living Systems (ExCELLS), National Institute of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, Japan; Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe-dori, Mizuhoku, Nagoya 467-8603, Japan; Institute for Molecular Science (IMS), National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, JapanJapan Collection of Microorganisms (JCM), RIKEN BioResource Research Center, Tsukuba, Ibaraki, JapanJapan Collection of Microorganisms (JCM), RIKEN BioResource Research Center, Tsukuba, Ibaraki, Japan; Submarine Resources Research Center, JAMSTEC, Yokosuka 273-0061, JapanJapan Collection of Microorganisms (JCM), RIKEN BioResource Research Center, Tsukuba, Ibaraki, JapanExploratory Research Center on Life and Living Systems (ExCELLS), National Institute of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, Japan; Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe-dori, Mizuhoku, Nagoya 467-8603, Japan; Institute for Molecular Science (IMS), National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, JapanInstitute for Extra-cutting-edge Science and Technology Avant-garde Research (X-star), Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima-cho, Yokosuka 273-0061, Japan; Exploratory Research Center on Life and Living Systems (ExCELLS), National Institute of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, JapanArchaeal cells are typically enveloped by glycosylated S-layer proteins. Archaeal protein glycosylation provides valuable insights not only into their adaptation to their niches but also into their evolutionary trajectory. Notably, thermophilic Thermoproteota modify proteins with N-glycans that include two GlcNAc units at the reducing end, resembling the ''core structure'' preserved across eukaryotes. Recently, Asgard archaea, now classified as members of the phylum Promethearchaeota, have offered unprecedented opportunities for understanding the role of archaea in eukaryogenesis. Despite the presence of genes indicative of protein N-glycosylation in this archaeal group, these have not been experimentally investigated. Here we performed a glycoproteome analysis of the firstly isolated Asgard archaeon Promethearchaeum syntrophicum. Over 700 different proteins were identified through high-resolution LC-MS/MS analysis, however, there was no evidence of either the presence or glycosylation of putative S-layer proteins. Instead, N-glycosylation in this archaeon was primarily observed in an extracellular solute-binding protein, possibly related to chemoreception or transmembrane transport of oligopeptides. The glycan modification occurred on an asparagine residue located within the conserved N-X-S/T sequon, consistent with the pattern found in other archaea, bacteria, and eukaryotes. Unexpectedly, three structurally different N-glycans lacking the conventional core structure were identified in this archaeon, presenting unique compositions that included atypical sugars. Notably, one of these sugars was likely HexNAc modified with a threonine residue, similar to modifications previously observed in mesophilic methanogens within the Methanobacteriati. Our findings advance our understanding of Asgard archaea physiology and evolutionary dynamics.http://www.sciencedirect.com/science/article/pii/S2667160324000061N-glycosylationAsgard archaeaGlycoproteomeEukaryogenesisS-layer
spellingShingle Satoshi Nakagawa
Hiroyuki Imachi
Shigeru Shimamura
Saeko Yanaka
Hirokazu Yagi
Maho Yagi-Utsumi
Hiroyuki Sakai
Shingo Kato
Moriya Ohkuma
Koichi Kato
Ken Takai
Characterization of protein glycosylation in an Asgard archaeon
BBA Advances
N-glycosylation
Asgard archaea
Glycoproteome
Eukaryogenesis
S-layer
title Characterization of protein glycosylation in an Asgard archaeon
title_full Characterization of protein glycosylation in an Asgard archaeon
title_fullStr Characterization of protein glycosylation in an Asgard archaeon
title_full_unstemmed Characterization of protein glycosylation in an Asgard archaeon
title_short Characterization of protein glycosylation in an Asgard archaeon
title_sort characterization of protein glycosylation in an asgard archaeon
topic N-glycosylation
Asgard archaea
Glycoproteome
Eukaryogenesis
S-layer
url http://www.sciencedirect.com/science/article/pii/S2667160324000061
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