Recombinant prespore-specific antigen from Dictyostelium discoideum is a β-sheet glycoprotein with a spacer peptide modified by O-linked N-acetylglucosamine

Natasha E. Zachara, Nicolle H. Packer, Mark D. Temple, Martin B. Slade, Daniel R. Jardine, Peter Karuso, Catherine J. Moss, Bridget C. Mabbutt, Paul M. G. Curmi, Keith L. Williams, Andrew A. Gooley

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Prespore-specific antigen (PsA) is a putative cell-adhesion molecule of the cellular slime mould Dictyostelium discoideum, which has a similar molecular architecture to several mammalian cell-surface proteins. It has an N-terminal globular domain presented to the extracellular environment on an O-glycosylated stem (glycopeptide) that is attached to the cell membrane through a glycosyl-PtdIns anchor. The sequence of PsA suggests that PsA may belong to a new family of cell-surface molecules and here we present information on the structure of the N-terminal globular domain and determine the reducing-terminal linkage of the O-glycosylation. To obtain a sufficient amount of pure protein, a secreted recombinant form of PsA (rPsA), was expressed in D. discoideum and characterised. 1H-NMR spectra of rPsA contained features consistent with a high degree of β-sheet in the N-terminal globular domain, a feature commonly observed in cell-adhesion proteins. Solid-phase Edman degradation of the glycopeptide of rPsA indicated that 14 of the 15 threonines and serines in the spacer region were glycosylated. The chemical structures of the O-glycosylations were determined to be single N-acetylglucosamine residues.

LanguageEnglish
Pages511-518
Number of pages8
JournalEuropean Journal of Biochemistry
Volume238
Issue number2
DOIs
Publication statusPublished - 1996

Fingerprint

Acetylglucosamine
Glycoproteins
Glycosylation
Antigens
Peptides
Dictyostelium
Glycopeptides
Dictyosteliida
Cell adhesion
Cell Adhesion Molecules
Threonine
Cell membranes
Phosphatidylinositols
Anchors
Cell Adhesion
Serine
Membrane Proteins
Proteins
Cell Membrane
Nuclear magnetic resonance

Keywords

  • cell-surface glycoproteins
  • dictyostelium discoideum
  • N-acetylglucosamine
  • O-glycosylation

Cite this

Zachara, Natasha E. ; Packer, Nicolle H. ; Temple, Mark D. ; Slade, Martin B. ; Jardine, Daniel R. ; Karuso, Peter ; Moss, Catherine J. ; Mabbutt, Bridget C. ; Curmi, Paul M. G. ; Williams, Keith L. ; Gooley, Andrew A. / Recombinant prespore-specific antigen from Dictyostelium discoideum is a β-sheet glycoprotein with a spacer peptide modified by O-linked N-acetylglucosamine. In: European Journal of Biochemistry. 1996 ; Vol. 238, No. 2. pp. 511-518.
@article{2a76ac90f318480581d98186db6ddb5e,
title = "Recombinant prespore-specific antigen from Dictyostelium discoideum is a β-sheet glycoprotein with a spacer peptide modified by O-linked N-acetylglucosamine",
abstract = "Prespore-specific antigen (PsA) is a putative cell-adhesion molecule of the cellular slime mould Dictyostelium discoideum, which has a similar molecular architecture to several mammalian cell-surface proteins. It has an N-terminal globular domain presented to the extracellular environment on an O-glycosylated stem (glycopeptide) that is attached to the cell membrane through a glycosyl-PtdIns anchor. The sequence of PsA suggests that PsA may belong to a new family of cell-surface molecules and here we present information on the structure of the N-terminal globular domain and determine the reducing-terminal linkage of the O-glycosylation. To obtain a sufficient amount of pure protein, a secreted recombinant form of PsA (rPsA), was expressed in D. discoideum and characterised. 1H-NMR spectra of rPsA contained features consistent with a high degree of β-sheet in the N-terminal globular domain, a feature commonly observed in cell-adhesion proteins. Solid-phase Edman degradation of the glycopeptide of rPsA indicated that 14 of the 15 threonines and serines in the spacer region were glycosylated. The chemical structures of the O-glycosylations were determined to be single N-acetylglucosamine residues.",
keywords = "cell-surface glycoproteins, dictyostelium discoideum, N-acetylglucosamine, O-glycosylation",
author = "Zachara, {Natasha E.} and Packer, {Nicolle H.} and Temple, {Mark D.} and Slade, {Martin B.} and Jardine, {Daniel R.} and Peter Karuso and Moss, {Catherine J.} and Mabbutt, {Bridget C.} and Curmi, {Paul M. G.} and Williams, {Keith L.} and Gooley, {Andrew A.}",
year = "1996",
doi = "10.1111/j.1432-1033.1996.0511z.x",
language = "English",
volume = "238",
pages = "511--518",
journal = "FEBS Journal",
issn = "1742-4658",
publisher = "Wiley-Blackwell, Wiley",
number = "2",

}

Recombinant prespore-specific antigen from Dictyostelium discoideum is a β-sheet glycoprotein with a spacer peptide modified by O-linked N-acetylglucosamine. / Zachara, Natasha E.; Packer, Nicolle H.; Temple, Mark D.; Slade, Martin B.; Jardine, Daniel R.; Karuso, Peter; Moss, Catherine J.; Mabbutt, Bridget C.; Curmi, Paul M. G.; Williams, Keith L.; Gooley, Andrew A.

In: European Journal of Biochemistry, Vol. 238, No. 2, 1996, p. 511-518.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Recombinant prespore-specific antigen from Dictyostelium discoideum is a β-sheet glycoprotein with a spacer peptide modified by O-linked N-acetylglucosamine

AU - Zachara, Natasha E.

AU - Packer, Nicolle H.

AU - Temple, Mark D.

AU - Slade, Martin B.

AU - Jardine, Daniel R.

AU - Karuso, Peter

AU - Moss, Catherine J.

AU - Mabbutt, Bridget C.

AU - Curmi, Paul M. G.

AU - Williams, Keith L.

AU - Gooley, Andrew A.

PY - 1996

Y1 - 1996

N2 - Prespore-specific antigen (PsA) is a putative cell-adhesion molecule of the cellular slime mould Dictyostelium discoideum, which has a similar molecular architecture to several mammalian cell-surface proteins. It has an N-terminal globular domain presented to the extracellular environment on an O-glycosylated stem (glycopeptide) that is attached to the cell membrane through a glycosyl-PtdIns anchor. The sequence of PsA suggests that PsA may belong to a new family of cell-surface molecules and here we present information on the structure of the N-terminal globular domain and determine the reducing-terminal linkage of the O-glycosylation. To obtain a sufficient amount of pure protein, a secreted recombinant form of PsA (rPsA), was expressed in D. discoideum and characterised. 1H-NMR spectra of rPsA contained features consistent with a high degree of β-sheet in the N-terminal globular domain, a feature commonly observed in cell-adhesion proteins. Solid-phase Edman degradation of the glycopeptide of rPsA indicated that 14 of the 15 threonines and serines in the spacer region were glycosylated. The chemical structures of the O-glycosylations were determined to be single N-acetylglucosamine residues.

AB - Prespore-specific antigen (PsA) is a putative cell-adhesion molecule of the cellular slime mould Dictyostelium discoideum, which has a similar molecular architecture to several mammalian cell-surface proteins. It has an N-terminal globular domain presented to the extracellular environment on an O-glycosylated stem (glycopeptide) that is attached to the cell membrane through a glycosyl-PtdIns anchor. The sequence of PsA suggests that PsA may belong to a new family of cell-surface molecules and here we present information on the structure of the N-terminal globular domain and determine the reducing-terminal linkage of the O-glycosylation. To obtain a sufficient amount of pure protein, a secreted recombinant form of PsA (rPsA), was expressed in D. discoideum and characterised. 1H-NMR spectra of rPsA contained features consistent with a high degree of β-sheet in the N-terminal globular domain, a feature commonly observed in cell-adhesion proteins. Solid-phase Edman degradation of the glycopeptide of rPsA indicated that 14 of the 15 threonines and serines in the spacer region were glycosylated. The chemical structures of the O-glycosylations were determined to be single N-acetylglucosamine residues.

KW - cell-surface glycoproteins

KW - dictyostelium discoideum

KW - N-acetylglucosamine

KW - O-glycosylation

UR - http://www.scopus.com/inward/record.url?scp=0029894645&partnerID=8YFLogxK

U2 - 10.1111/j.1432-1033.1996.0511z.x

DO - 10.1111/j.1432-1033.1996.0511z.x

M3 - Article

VL - 238

SP - 511

EP - 518

JO - FEBS Journal

T2 - FEBS Journal

JF - FEBS Journal

SN - 1742-4658

IS - 2

ER -