Crystallization and Preliminary X-ray Diffraction Characterisation of Both a Native and Selenomethionyl VLA-4 Binding Fragment of VCAM-1

• Published in: Journal of molecular biology Vol. 244; no. 4; pp. 464 - 468
• Main Authors: 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.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 09.12.1994
• Subjects: adhesion molecule; Cell Adhesion Molecules - chemistry; Crystallography, X-Ray; integrin binding; Integrins - metabolism; Selenomethionine - chemistry; Vascular Cell Adhesion Molecule-1; VCAM-1; X-ray crystallography, selenomethionine; X-Ray Diffraction; VCAM-1; X-ray crystallography, selenomethionine; integrin binding; adhesion molecule
• ISSN: 0022-2836; 1089-8638
• DOI: 10.1006/jmbi.1994.1743

Soluble fragments of the extracellular region of vascular cell adhesion molecule 1 (VCAM-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 P 2 12 12 1 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 selenomethionyl crystals diffract X-rays to a minimum Bragg spacing of 1.8 Å