A rationally engineered yeast pyruvyltransferase Pvg1p introduces sialylation-like properties in neo-human-Type complex oligosaccharide

Yujiro Higuchi, Sho Yoshinaga, Ken Ichi Yoritsune, Hiroaki Tateno, Jun Hirabayashi, Shin Ichi Nakakita, Miho Kanekiyo, Yoshimitsu Kakuta, Kaoru Takegawa

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Pyruvylation onto the terminus of oligosaccharide, widely seen from prokaryote to eukaryote, confers negative charges on the cell surface and seems to be functionally similar to sialylation, which is found at the end of human-Type complex oligosaccharide. However, detailed molecular mechanisms underlying pyruvylation have not been clarified well. Here, we first determined the crystal structure of fission yeast pyruvyltransferase Pvg1p at a resolution of 2.46 Å. Subsequently, by combining molecular modeling with mutational analysis of active site residues, we obtained a Pvg1p mutant (Pvg1p H168C) that efficiently transferred pyruvyl moiety onto a human-Type complex glycopeptide. The resultant pyruvylated human-Type complex glycopeptide recognized similar lectins on lectin arrays as the α2,6-sialyl glycopeptides. This newly-generated pyruvylation of human-Type complex oligosaccharides would provide a novel method for glyco-bioengineering.

Original languageEnglish
Article number26349
JournalScientific reports
Volume6
DOIs
Publication statusPublished - May 19 2016

Fingerprint

Oligosaccharides
Glycopeptides
Yeasts
Lectins
Bioengineering
Schizosaccharomyces
Eukaryota
Catalytic Domain

All Science Journal Classification (ASJC) codes

  • General

Cite this

A rationally engineered yeast pyruvyltransferase Pvg1p introduces sialylation-like properties in neo-human-Type complex oligosaccharide. / Higuchi, Yujiro; Yoshinaga, Sho; Yoritsune, Ken Ichi; Tateno, Hiroaki; Hirabayashi, Jun; Nakakita, Shin Ichi; Kanekiyo, Miho; Kakuta, Yoshimitsu; Takegawa, Kaoru.

In: Scientific reports, Vol. 6, 26349, 19.05.2016.

Research output: Contribution to journalArticle

Higuchi, Yujiro ; Yoshinaga, Sho ; Yoritsune, Ken Ichi ; Tateno, Hiroaki ; Hirabayashi, Jun ; Nakakita, Shin Ichi ; Kanekiyo, Miho ; Kakuta, Yoshimitsu ; Takegawa, Kaoru. / A rationally engineered yeast pyruvyltransferase Pvg1p introduces sialylation-like properties in neo-human-Type complex oligosaccharide. In: Scientific reports. 2016 ; Vol. 6.
@article{000a5951deb543af8ba787f97a9cc218,
title = "A rationally engineered yeast pyruvyltransferase Pvg1p introduces sialylation-like properties in neo-human-Type complex oligosaccharide",
abstract = "Pyruvylation onto the terminus of oligosaccharide, widely seen from prokaryote to eukaryote, confers negative charges on the cell surface and seems to be functionally similar to sialylation, which is found at the end of human-Type complex oligosaccharide. However, detailed molecular mechanisms underlying pyruvylation have not been clarified well. Here, we first determined the crystal structure of fission yeast pyruvyltransferase Pvg1p at a resolution of 2.46 {\AA}. Subsequently, by combining molecular modeling with mutational analysis of active site residues, we obtained a Pvg1p mutant (Pvg1p H168C) that efficiently transferred pyruvyl moiety onto a human-Type complex glycopeptide. The resultant pyruvylated human-Type complex glycopeptide recognized similar lectins on lectin arrays as the α2,6-sialyl glycopeptides. This newly-generated pyruvylation of human-Type complex oligosaccharides would provide a novel method for glyco-bioengineering.",
author = "Yujiro Higuchi and Sho Yoshinaga and Yoritsune, {Ken Ichi} and Hiroaki Tateno and Jun Hirabayashi and Nakakita, {Shin Ichi} and Miho Kanekiyo and Yoshimitsu Kakuta and Kaoru Takegawa",
year = "2016",
month = "5",
day = "19",
doi = "10.1038/srep26349",
language = "English",
volume = "6",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",

}

TY - JOUR

T1 - A rationally engineered yeast pyruvyltransferase Pvg1p introduces sialylation-like properties in neo-human-Type complex oligosaccharide

AU - Higuchi, Yujiro

AU - Yoshinaga, Sho

AU - Yoritsune, Ken Ichi

AU - Tateno, Hiroaki

AU - Hirabayashi, Jun

AU - Nakakita, Shin Ichi

AU - Kanekiyo, Miho

AU - Kakuta, Yoshimitsu

AU - Takegawa, Kaoru

PY - 2016/5/19

Y1 - 2016/5/19

N2 - Pyruvylation onto the terminus of oligosaccharide, widely seen from prokaryote to eukaryote, confers negative charges on the cell surface and seems to be functionally similar to sialylation, which is found at the end of human-Type complex oligosaccharide. However, detailed molecular mechanisms underlying pyruvylation have not been clarified well. Here, we first determined the crystal structure of fission yeast pyruvyltransferase Pvg1p at a resolution of 2.46 Å. Subsequently, by combining molecular modeling with mutational analysis of active site residues, we obtained a Pvg1p mutant (Pvg1p H168C) that efficiently transferred pyruvyl moiety onto a human-Type complex glycopeptide. The resultant pyruvylated human-Type complex glycopeptide recognized similar lectins on lectin arrays as the α2,6-sialyl glycopeptides. This newly-generated pyruvylation of human-Type complex oligosaccharides would provide a novel method for glyco-bioengineering.

AB - Pyruvylation onto the terminus of oligosaccharide, widely seen from prokaryote to eukaryote, confers negative charges on the cell surface and seems to be functionally similar to sialylation, which is found at the end of human-Type complex oligosaccharide. However, detailed molecular mechanisms underlying pyruvylation have not been clarified well. Here, we first determined the crystal structure of fission yeast pyruvyltransferase Pvg1p at a resolution of 2.46 Å. Subsequently, by combining molecular modeling with mutational analysis of active site residues, we obtained a Pvg1p mutant (Pvg1p H168C) that efficiently transferred pyruvyl moiety onto a human-Type complex glycopeptide. The resultant pyruvylated human-Type complex glycopeptide recognized similar lectins on lectin arrays as the α2,6-sialyl glycopeptides. This newly-generated pyruvylation of human-Type complex oligosaccharides would provide a novel method for glyco-bioengineering.

UR - http://www.scopus.com/inward/record.url?scp=84970016142&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84970016142&partnerID=8YFLogxK

U2 - 10.1038/srep26349

DO - 10.1038/srep26349

M3 - Article

VL - 6

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

M1 - 26349

ER -