Abstract
A putative mature human neutrophil chemotactic factor (NCF) corresponding to the C-terminal 72 amino acids of its precursor was directly produced in Escherichia coli by recombinant DNA technology. Human NCF was present in both the soluble and insoluble protein fractions of the homogenate of host cells, and it was partially purified as a water-soluble polypeptide from both fractions, separately. The partially purified NCF preparation was highly purified to an endotoxin-free homogeneous polypeptide by means of CM-Sepharose CL-6B column chromatography and gel filtration on Toyopearl HW-55. No difference between the human NCF preparations purified from both starting materials could be found concerning purity, primary structure, solubility, molecular weight, and chemotactic activity for human neutrophils. The amino acid sequence of recombinant human NCF was identical to the sequence deduced from the cDNA sequence. A methionine residue due to the translation initiation codon was removed. Recombinant human NCF was found to be biologically active and to exhibit chemotactic activity for human neutrophils in vitro and cause a neutrophil infiltration in vitro in mice.
| Original language | English |
|---|---|
| Pages (from-to) | 436-441 |
| Number of pages | 6 |
| Journal | Journal of Biochemistry |
| Volume | 106 |
| Issue number | 3 |
| Publication status | Published - Sep 1989 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Statistics, Probability and Uncertainty
- Applied Mathematics
- Physiology (medical)
- Radiology Nuclear Medicine and imaging
- Molecular Biology
- Biochemistry
Cite this
Production and characterization of recombinant human neutrophil chemotactic factor. / Furuta, Ryuji; Yamagishi, Junichi; Kotani, Hirotada; Sakamoto, Fumiko; Fukui, Toshikazu; Matsui, Yukiharu; Sohmura, Yasunobu; Yamada, Masaaki; Yoshimura, Teizo; Larsen, Christian G.; Oppenheim, Joost J.; Matsushima, Kouji.
In: Journal of Biochemistry, Vol. 106, No. 3, 09.1989, p. 436-441.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Production and characterization of recombinant human neutrophil chemotactic factor
AU - Furuta, Ryuji
AU - Yamagishi, Junichi
AU - Kotani, Hirotada
AU - Sakamoto, Fumiko
AU - Fukui, Toshikazu
AU - Matsui, Yukiharu
AU - Sohmura, Yasunobu
AU - Yamada, Masaaki
AU - Yoshimura, Teizo
AU - Larsen, Christian G.
AU - Oppenheim, Joost J.
AU - Matsushima, Kouji
PY - 1989/9
Y1 - 1989/9
N2 - A putative mature human neutrophil chemotactic factor (NCF) corresponding to the C-terminal 72 amino acids of its precursor was directly produced in Escherichia coli by recombinant DNA technology. Human NCF was present in both the soluble and insoluble protein fractions of the homogenate of host cells, and it was partially purified as a water-soluble polypeptide from both fractions, separately. The partially purified NCF preparation was highly purified to an endotoxin-free homogeneous polypeptide by means of CM-Sepharose CL-6B column chromatography and gel filtration on Toyopearl HW-55. No difference between the human NCF preparations purified from both starting materials could be found concerning purity, primary structure, solubility, molecular weight, and chemotactic activity for human neutrophils. The amino acid sequence of recombinant human NCF was identical to the sequence deduced from the cDNA sequence. A methionine residue due to the translation initiation codon was removed. Recombinant human NCF was found to be biologically active and to exhibit chemotactic activity for human neutrophils in vitro and cause a neutrophil infiltration in vitro in mice.
AB - A putative mature human neutrophil chemotactic factor (NCF) corresponding to the C-terminal 72 amino acids of its precursor was directly produced in Escherichia coli by recombinant DNA technology. Human NCF was present in both the soluble and insoluble protein fractions of the homogenate of host cells, and it was partially purified as a water-soluble polypeptide from both fractions, separately. The partially purified NCF preparation was highly purified to an endotoxin-free homogeneous polypeptide by means of CM-Sepharose CL-6B column chromatography and gel filtration on Toyopearl HW-55. No difference between the human NCF preparations purified from both starting materials could be found concerning purity, primary structure, solubility, molecular weight, and chemotactic activity for human neutrophils. The amino acid sequence of recombinant human NCF was identical to the sequence deduced from the cDNA sequence. A methionine residue due to the translation initiation codon was removed. Recombinant human NCF was found to be biologically active and to exhibit chemotactic activity for human neutrophils in vitro and cause a neutrophil infiltration in vitro in mice.
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UR - http://www.scopus.com/inward/citedby.url?scp=0024425808&partnerID=8YFLogxK
M3 - Article
C2 - 2691501
AN - SCOPUS:0024425808
VL - 106
SP - 436
EP - 441
JO - Journal of Biochemistry
JF - Journal of Biochemistry
SN - 0021-924X
IS - 3
ER -