Expression, crystallization, and preliminary X-ray analysis of a sialic acid-binding fragment of sialoadhesin in the presence and absence of ligand

A. P. May, R. C. Robinson, R. T. Aplin, P. Bradfield, P. R. Crocker, E. Y. Jones

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Sialoadhesin is a macrophage-restricted cell surface receptor, consisting of 17 immunoglobulin domains, which mediates cell adhesion via the recognition of specific sialylated glycoconjugates. A functional fragment of sialoadhesin, comprising the N-terminal immunoglobulin domain, has been expressed in Chinese hamster ovary cells as both native (SnD1) and selenomethionyl (Se-SnD1) stop protein. The successful production of 86% selenomethionine-incorporated protein represents a rare example of production of selenium-labeled protein in mammalian cells. SnD1 and Se-SnD1 have been crystallized in the absence of ligand, and SnD1 has also been crystallized in the presence of its ligand 2,3 sialyllactose. The ligand-free crystals of SnD1 and Se-SnD1 were isomorphous, of space group P3121 or P3221, with unit cell dimensions a = b = 38.9 Å, c = 152.6 Å, α = β = 90°, γ 120°, and diffracted to a maximum resolution of 2.6 Å. Cocrystals containing 2,3 sialyllactose diffracted to 1.85 Å at a synchrotron source and belong to space group P212121, with unit cell dimensions α = 40.9 Å, b = 97.6 Å, c = 101.6 Å, α = β = υ = 90°.

Original languageEnglish
Pages (from-to)717-721
Number of pages5
JournalProtein Science
Volume6
Issue number3
Publication statusPublished - Mar 1 1997
Externally publishedYes

Fingerprint

Sialic Acid Binding Ig-like Lectin 1
X ray analysis
N-Acetylneuraminic Acid
Crystallization
X-Rays
Ligands
Immunoglobulins
Cells
Selenomethionine
Proteins
Glycoconjugates
Macrophages
Cell adhesion
Cell Surface Receptors
Selenium
Synchrotrons
Cricetulus
Cell Adhesion
Ovary
Crystals

Keywords

  • cell adhesion
  • complex
  • crystallization
  • lectin
  • selenomethionine
  • sialic acid

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Expression, crystallization, and preliminary X-ray analysis of a sialic acid-binding fragment of sialoadhesin in the presence and absence of ligand. / May, A. P.; Robinson, R. C.; Aplin, R. T.; Bradfield, P.; Crocker, P. R.; Jones, E. Y.

In: Protein Science, Vol. 6, No. 3, 01.03.1997, p. 717-721.

Research output: Contribution to journalArticle

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