Production and characterization of recombinant human neutrophil chemotactic factor.
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 vivo in mice.
Furuta
R
R
Research Laboratories, Dainippon Pharmaceutical Co., Ltd., Osaka.
Yamagishi
J
J
Kotani
H
H
Sakamoto
F
F
Fukui
T
T
Matsui
Y
Y
Sohmura
Y
Y
Yamada
M
M
Yoshimura
T
T
Larsen
C G
CG
D000595:Amino Acid Sequence; D000818:Animals; D002465:Cell Movement; D002630:Chemotactic Factors / Q000096:biosynthesis* / Q000302:isolation & purification / Q000502:physiology; D002634:Chemotaxis, Leukocyte; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D016209:Interleukin-8; D051379:Mice; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000502:physiology; D011994:Recombinant Proteins / Q000096:biosynthesis*
D002630:Chemotactic Factors; D016209:Interleukin-8; D011994:Recombinant Proteins