Crystallization and preliminary X-ray diffraction characterisation of both a native and selenomethionyl VLA-4 binding fragment of VCAM-1

M. J. Bottomley, R. C. Robinson, P. C. Driscoll, K. Harlos, D. I. Stuart, R. T. Aplin, J. M. Clements, E. Y. Jones, T. J. Dudgeon

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8 Citations (Scopus)

Abstract

Soluble fragments of the extracellular region of vascular cell adhesion molecule 1 (VCBM-1) expressed in Escherichia coli retain functional adhesive activity. An integrin (VLA-4) binding fragment consisting of the N-terminal two immunoglobulin-like domains (VCAM-d1,2) has been crystallized. The crystals belong to space group P212121 with cell dimensions of a = 52.7 Å, b = 66.5 Å, c = 113.2 Å and contain two molecules in the crystallographic asymmetric unit. A batch of protein produced in the standard E. coli strain (HW1110), but grown in the presence of selenomethionine enriched media, showed 85% incorporation of selenium in place of sulphur at methionine residues. The selenomethionyl VCAM-d1,2 was crystallized by microseeding techniques initially using the native crystals for nucleation. Both native and selenomethiongrl crystals diffract X-rays to a minimum Bragg spacing of 1.8 Å.

Original languageEnglish
Pages (from-to)464-468
Number of pages5
JournalJournal of Molecular Biology
Volume244
Issue number4
DOIs
Publication statusPublished - Dec 8 1994
Externally publishedYes

Keywords

  • Adhesion molecule
  • Integrin binding
  • Selenomethionine
  • VCAM-1
  • X-ray crystallography

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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    Bottomley, M. J., Robinson, R. C., Driscoll, P. C., Harlos, K., Stuart, D. I., Aplin, R. T., Clements, J. M., Jones, E. Y., & Dudgeon, T. J. (1994). Crystallization and preliminary X-ray diffraction characterisation of both a native and selenomethionyl VLA-4 binding fragment of VCAM-1. Journal of Molecular Biology, 244(4), 464-468. https://doi.org/10.1006/jmbi.1994.1743