Crystallization of trimeric recombinant human tumor necrosis factor (cachectin)

Authors

M. J. Eck, University of Texas Southwestern Medical Center
B. Beutler, University of Texas Southwestern Medical Center
G. Kuo, University of Texas Southwestern Medical Center
J. P. Merryweather, University of Texas Southwestern Medical Center
S. R. Sprang, University of Montana

Document Type

Article

Publication Title

Journal of Biological Chemistry

Publication Date

9-1988

Volume

263

Issue

26

Disciplines

Biology | Life Sciences

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.

Rights

© 1988 by The American Society for Biochemistry and Molecular Biology, Inc.

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Recommended Citation

Eck, M. J.; Beutler, B.; Kuo, G.; Merryweather, J. P.; and Sprang, S. R., "Crystallization of trimeric recombinant human tumor necrosis factor (cachectin)" (1988). Biological Sciences Faculty Publications. 559.
https://scholarworks.umt.edu/biosci_pubs/559