title: Crystallization of purified recombinant human interleukin-1 beta.
abstract: The gene for human interleukin-1 beta was cloned from SK-hep-1 hepatoma cellular RNA and expressed at high levels in Escherichia coli both as the naturally processed form (rIL-1 beta) and as a variant with an additional sequence of three amino acids on the N-terminus (rIL-1 beta +). Expressed protein was purified to homogeneity by a sequence of steps, which included low pH incubation, adsorption and desorption from Procion Red Sepharose, sizing on a Superose 12 fast-performance liquid chromatography (FPLC) column, and anion exchange chromatography on QAE Sepharose. The final step provided a biologically active protein that migrates on two-dimensional (2-D) gels as a single spot with a pI of 6.7 +/- 0.2 and a molecular mass of 17,500 daltons. Concentrated solutions of rIL-1 beta have produced crystals by ammonium sulfate precipitation. The crystals are tetragonal, show the symmetry of space group P4(1) or its enantiomer, have lattice constants of a = 58.46 (1) and c = 77.02 (3) A, and scatter to at least 2 A resolution. A structure determination based on these crystals is under way.
authors.affiliation: Molecular Biology Research, Upjohn Company, Kalamazoo, Michigan 49007.
authors.forename: D B
authors.initials: DB
authors.lastname: Carter
authors.forename: K A
authors.initials: KA
authors.lastname: Curry
authors.forename: C S
authors.initials: CS
authors.lastname: Tomich
authors.forename: A W
authors.initials: AW
authors.lastname: Yem
authors.forename: M R
authors.initials: MR
authors.lastname: Deibel
authors.forename: D E
authors.initials: DE
authors.lastname: Tracey
authors.forename: J W
authors.initials: JW
authors.lastname: Paslay
authors.forename: J B
authors.initials: JB
authors.lastname: Carter
authors.forename: N Y
authors.initials: NY
authors.lastname: Theriault
authors.forename: P K
authors.initials: PK
authors.lastname: Harris
mesh_terms: D000595:Amino Acid Sequence; D006528:Carcinoma, Hepatocellular; D002460:Cell Line; D003001:Cloning, Molecular; D003460:Crystallization; D004262:DNA Restriction Enzymes; D005796:Genes; D006801:Humans; D007375:Interleukin-1 / Q000235:genetics / Q000302:isolation & purification*; D008113:Liver Neoplasms; D010957:Plasmids; D011994:Recombinant Proteins / Q000302:isolation & purification*
chemical_list: D007375:Interleukin-1; D011994:Recombinant Proteins; D004262:DNA Restriction Enzymes