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

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...

Full description

Saved in:
Bibliographic Details
Published in:FEBS letters 1994-06, Vol.347 (2), p.300-303
Main Authors: Kungl, Andreas J., Machius, Mischa, Huber, Robert, Schwer, Christine, Lam, Charles, Aschauer, Heinz, Ehn, Gerald, Lindley, Ivan J.D., Auer, Manfred
Format: Article
Language:English
Subjects:
aa, amino acid
Amino Acid Sequence
AS, ammonium sulfate
beta-Thromboglobulin
Chemokine
Chromatography, High Pressure Liquid
Cloning, Molecular
Crystallization
Crystallography, X-Ray
Cytokine
Electrophoresis, Polyacrylamide Gel
Escherichia coli - genetics
FPLC, fast performance liquid chromatography
Gene Expression
GuHCl, guanidinium hydrochloride
HPLC, high pressure liquid chromatography
Humans
Leukocyte Elastase
MES, morpholino ethane sulfonic acid
Molecular Sequence Data
NAP-1/IL-8, neutrophil-activating peptide 1/interleukin 8
Neutrophil-activating peptide 2
Neutrophils - drug effects
Neutrophils - enzymology
Pancreatic Elastase - metabolism
PEG, polyethylene glycol
Peptides - chemistry
Peptides - isolation & purification
Peptides - pharmacology
Recombinant Proteins - chemistry
Recombinant Proteins - isolation & purification
rhNAP-2, recombinant human neutrophil-activating peptide 2
X-ray diffraction
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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 Å 3/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.
ISSN:0014-5793
1873-3468
DOI:10.1016/0014-5793(94)00573-7