Crystallization of trimeric recombinant human tumor necrosis factor (cachectin)

M. J. Eck, B. Beutler, G. Kuo, J. P. Merryweather, S. R. Sprang

Research output: Contribution to journalArticle

37 Scopus citations

Abstract

Crystals of tumor necrosis factor (TNF) have been obtained in two forms. Rhombohedral crystals grow in 1.8 to 2.0 M ammonium sulfite, pH 7.8 at 21°C, and tetragonal crystals grow in 2.6 M magnesium sulfate, pH 5.5 at 25°C. Analysis of TNF by isoelectric focusing under native and denaturing conditions indicates that TNF molecules exist as trimers in solution. The rhombohedral cachectin crystals belong to space group R3 and have unit cell constants a = b = c = 47.65 Å and α = β = γ = 88.1°. Density determinations and the space group indicate that the unit cell contains one 51,000-dalton trimer. These crystals are stable in the x-ray beam and diffract to at least 1.85 Å but are apparently twinned by merohedry. The tetragonal crystals are space group P43212 or its enantiomorph P41212 and have unit cell constants a = b = 95.08, c = 117.49. The asymmetric unit contains one trimer; the crystals are stable in the x-ray beam and diffract to beyond 3 Å.

Original languageEnglish (US)
Pages (from-to)12816-12819
Number of pages4
JournalJournal of Biological Chemistry
Volume263
Issue number26
StatePublished - 1988

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Eck, M. J., Beutler, B., Kuo, G., Merryweather, J. P., & Sprang, S. R. (1988). Crystallization of trimeric recombinant human tumor necrosis factor (cachectin). Journal of Biological Chemistry, 263(26), 12816-12819.