Crystallization of recombinant human interleukin 1β

• Published in: Journal of crystal growth 1988-07, Vol.90 (1-3), p.180-187
• Main Authors: Einspahr, Howard, Clancy, L.L., Muchmore, S.W., Watenpaugh, K.D., Harris, P.K.W., Carter, D.B., Curry, K.A., Tomich, C.-S.C., Yem, A.W., Deibel, M.R., Tracey, D.E., Paslay, J.W., Staite, N.D., Carter, J.B., Theriault, N.Y., Reardon, I.M., Zurcher-Neely, H.A., Heinrikson, R.L.
• Format: Article
• Language: English
• ISSN: 0022-0248; 1873-5002
• DOI: 10.1016/0022-0248(88)90313-2

The gene for the fully processed form of human interleukin 1β was cloned from SK-hep-1 hepatoma cellular RNA and expressed at high levels in E. coli. The protein produced in E. coli. was purified to homogeneity by standard chromatographic methods, including adsorption and desorption from Procion Red Sepharose, sizing on a Superose 12 FPLC column, and anion exchange chromatography on QAE Sepharose. The result is a biologically active protein, rIL-1β, that migrates on two-dimensional gels as a single spot with a pI of 6.5 ± 0.2 and a molecular mass of 17, 500 daltons. Crystals of rIL-1β have been produced from concentrated solutions of the protein by ammonium sulfate precipitation. The crystals are tetragonal, have space group P41 or its enantiomer, have lattice constants of a = 58.46(1) Å and c = 77.02(3) Å, and scatter to at least 2 Å resolution. A structure determination ba these crystals is underway.