Crystallization and preliminary X‐ray crystallographic study of interleukin‐8

Auer, Manfred; Kallen, Joerg; Schleischitz, Sabine; Walkinshaw, Malcolm D.; Wasserbauer, Erich; Ehn, Gerald; Lindley, I. J. D.

doi:10.1016/0014-5793(90)80876-k

Cited by 6 publications

(2 citation statements)

References 18 publications

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“…Identical crystals have also been grown using chemically synthesized protein. Recently, workers at Sandoz Pharmaceutical have reported that crystals apparently identical to the ones described here could be grown from polyethylene glycol at pH 6.5 (20). The crystals contain a monomer in the asymmetric unit.…”

Section: Methodssupporting

confidence: 67%

Crystal structure of interleukin 8: symbiosis of NMR and crystallography.

Baldwin

Weber

Charles

et al. 1991

Proc. Natl. Acad. Sci. U.S.A.

339

223

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The crystal structure of a host defense system chemotactic factor, interleukin 8, has been solved by molecular replacement using as a model the solution structure derived from nuclear magnetic resonance experiments. The structure was refined with 2 A x-ray data to an R factor of 0.187 (0.217 at 1.6 A). A comparison indicates some potential differences between the structure in solution and in the crystalline state. Our analysis also predicts that residues 4 through 9 on the amino terminus and the (-bend, which includes His-33, may be important for receptor binding.Nuclear magnetic resonance (NMR) has been established in the last 5 years as the only viable alternative to x-ray crystallography for determining detailed three-dimensional structures of macromolecules up to 20 kDa molecular mass (1, 2). Although test calculations using crambin (3), tendamistat (4), and ubiquitin (5) have established the feasibility of utilizing NMR-derived models as starting points for solving x-ray structures by molecular replacement, no unknown crystal structures have been previously solved by that route. We present here the results of such an investigation. The highresolution x-ray structure ofinterleukin 8 (IL-8) was solved by a combination of NMR (6, 7) and crystallography, and the models obtained by these two techniques were compared.tt IL-8 is a small, chemotactic factor released by monocytes and a number of other cell types in response to inflammatory stimuli (8). The factor promotes the directed migration of neutrophils, basophils, and T cells (9) along a concentration gradient to the inflammatory focus. IL-8 has also been reported to initiate the oxidative burst response in neutrophils. The factor has been known as monocyte-derived neutrophil chemotactic factor (10) or neutrophil-activating protein (11,12) among others, but it is now generally designated IL-8 (9). IL-8 is a member of a family of similar immune system proteins including the monocyte chemoattractant factor (13), human melanoma growth-stimulating activity (14), murine macrophage inflammatory protein 2 (15), and bovine platelet factor 4 (BPF4) (16). Each of these protein sequences shows a conserved region containing four cysteines that form two disulfide bridges. Under the experimental conditions of the structural investigations IL-8 is a dimer (6, 7) and BPF4 is a tetramer (17). Materials and MethodsRecombinant human IL-8 that was used in the crystallographic studies was purified from Escherichia coli (18). All of the x-ray data used for this structure determination were collected on crystals that were grown using protein recycled from the NMR experiments of Clore and Gronenborn and coworkers (6, 7). Crystals belonging to the space group P3121, a = b = 40.3 A, c = 90.1 A, were grownfrom40%o ammonium sulfate (pH = 8.5) as described (19). Identical crystals have also been grown using chemically synthesized protein. Recently, workers at Sandoz Pharmaceutical have reported that crystals apparently identical to the ones described here could be grown from poly...

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Section: Methodssupporting

confidence: 67%

Crystal structure of interleukin 8: symbiosis of NMR and crystallography.

Baldwin

Weber

Charles

et al. 1991

Proc. Natl. Acad. Sci. U.S.A.

339

223

View full text Add to dashboard Cite

show abstract

“…This cytokine was termed interleukin-8 (IL-8) [29]. Subsequently, it has been shown that many cell types other than monocytes, including blood monocytes, alveolar macrophages, fibroblasts, endothelial cells, and epithelial cells, can produce large quantities of IL-8 or IL-8 mRNA, in response to stimulation with endotoxin, IL-l, or tumor necrosis factor [8,30]. The functional study revealed that IL-8 overexpressed on some tumor cells, and that tumor-derived IL-8 through autocrine and paracrine could alter the composition of immune infiltrates in the tumor microenvironment and induce angiogenesis and growth of the cancer cells [31], which even involved in EMT and CSC-like change.…”

Section: Discussionmentioning

confidence: 99%