Purification, crystallization and preliminary X‐ray diffraction analysis of recombinant human neutrophil‐activating peptide 2 (rhNAP‐2)

FEBS Letters

Auer, Manfred¹  Lam, Charles¹  Schwer, Christine²  Aschauer, Heinz¹  Kungl, Andreas J.²  Huber, Robert²  Lindley, Ivan J.D.¹  Ehn, Gerald¹  Machius, Mischa² 

[1] Sandoz Research Institute, Brunnerstraße 59, A-1235 Wien, Austria;Max-Planck-Institut für Biochemie, Am Klopferspitz 18a, D-82152 Martinsried, Germany

关键词: Neutrophil-activating peptide 2;    Chemokine;    Cytokine;    Crystallization;    X-ray diffraction;    rhNAP-2;    recombinant human neutrophil-activating peptide 2;    NAP-1/IL-8;    neutrophil-activating peptide 1/interleukin 8;    HPLC;    high pressure liquid chromatography;    GuHCl;    guanidinium hydrochloride;    MES;    morpholino ethane sulfonic acid;    FPLC;    fast performance liquid chromatography;    PEG;    polyethylene glycol;    AS;    ammonium sulfate;    aa;    amino acid;   

DOI  :  10.1016/0014-5793(94)00573-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The potent activator and chemoattractant for human neutrophils, neutrophil-activating peptide 2 (NAP-2), has been cloned and expressed in Escherichia coli. The protein has been purified to homogeneity (> 98%) by a series of chromatographic techniques, including reversed phase HPLC. The biological activity of recombinant human NAP-2 (rhNAP-2), characterized by the induction of elastase release from human neutrophils, was found to be comparable to natural NAP-2. rhNAP-2 has been crystallized by the hanging drop vapor diffusion method. The crystals belong to space group P222 with unit cell dimensions of a = 30.8 Å, b = 39.5 Å and c = 95.3 Å. A packing density of 3.8 ų/Da with a solvent content of approximately 68% is obtained when one molecule per asymmetric unit is assumed. The crystals were shown to diffract to beyond 2.0 Å on a conventional X-ray source. They are stable to X-rays for several days and are thus suitable for high resolution structure determination.

【 授权许可】

Unknown