Purification and amino acid sequencing of NAF, a novel neutrophil-activating factor produced by monocytes.
Human blood mononuclear cells were cultured for 24 h in the presence of LPS (100 ng per 5 x 10(6) cells), and a monocyte-derived neutrophil-activating factor (NAF) was purified to apparent homogeneity from the conditioned media. The purification consisted of ammonium sulphate precipitation, gel filtration, chromatography on phosphocellulose followed by hydroxylapatite, and reversed-phase HPLC on C4 and CN-propyl columns. Amino acid sequence analysis (32 of 50 presumed residues) shows that NAF is a novel peptide with little homology to known ones. Crude and pure NAF stimulated human neutrophils to release granule enzymes and to produce superoxide and H2O2.
Walz
A
A
Theodor-Kocher-Institut, University of Bern, Switzerland.
Peveri
P
P
Aschauer
H
H
Baggiolini
M
M
D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D016209:Interleukin-8; D008969:Molecular Sequence Data; D009000:Monocytes / Q000378:metabolism*; D009504:Neutrophils / Q000187:drug effects* / Q000378:metabolism; D010455:Peptides / Q000032:analysis / Q000302:isolation & purification*
D016209:Interleukin-8; D010455:Peptides