Crystallization and preliminary X-ray investigation reveals that tumor necrosis factor is a compact trimer furnished with 3-fold symmetry

Abstract

Crystals of recombinant human tumor necrosis factor produced by Escherichia coli have been obtained under different conditions. Crystals suitable for X-ray studies are produced by a vapor diffusion technique using sodium phosphate as both precipitant and buffer at pH 6·5. The crystals belong to the cubic space group, P2 13 with unit cell dimensions a = b = c = 95·7 A ̊ (1 Å = 0·1 nm). Preliminary photography reveals that the crystals are moderately stable to X-rays and diffract to at least 3 Å resolution. The diffraction data for native crystals have been collected on a diffractometer at 3 Å resolution. Another crystal form, which appeared in a solution containing sodium phosphate at pH 8·0, has the trigonal space group P3 with unit cell dimensions a = b = 63·8 A ̊ and c = 54·4 A ̊ , and produces measurable reflections to a resolution of 3 Å. Hexagonal crystals also have been obtained by the use of polyethylene glycol as precipitant in the range pH 7·6 to 8·0; however, the crystals are fragile and unstable to X-rays. Conservation of 3-fold symmetry in the different crystal forms obtained could reflect the ability of tumor necrosis factor molecules to form trimers in solution and probably the nature of binding of the molecules to cellular receptors.