Purification and partial primary sequence of a chemotactic protein for polymorphonuclear leukocytes derived from human lung giant cell carcinoma LU65C cells.
A chemotactic protein for polymorphonuclear leukocytes (lung carcinoma-derived chemotaxin [LUCT]) was purified from culture fluid of the human lung giant cell carcinoma LU65C cells to electrophoretically homogeneous form through five sequential purification steps: DEAE-Sepharose, CM-Sepharose, HPLC on carboxyl-methylated-polyvinylalcohol resin, hydrophobic, and reversed-phase. The molecular mass was determined as approximately 10 kD by SDS-PAGE and isoelectric point was 10.7. The chemotactic activity (ED50 0.75 x 10(-9) M) was sevenfold more potent than that of FMLP (5 X 10(-9) M) and comparable with that of C5a (10(-9) M). NH2-terminal amino acid sequence and amino acid composition of LUCT strongly suggest that it may be closely related to the putative protein encoded by the cDNA clone (3-10C) and almost identical with a part of sequence of the chemotactic factor derived from stimulated human leukocytes in the 6th to 32nd, but not the NH2-terminal 5 amino acids. These results indicate that the carcinoma cells produce LUCT without any added stimulant and suggest that the previously isolated chemotactic monokines may correspond to des(1-5) of LUCT in the NH2-terminal region.
Suzuki
K
K
Department of Antibiotics, National Institute of Health, Tokyo, Japan.
Miyasaka
H
H
Ota
H
H
Yamakawa
Y
Y
Tagawa
M
M
Kuramoto
A
A
Mizuno
S
S
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000302:isolation & purification; D002277:Carcinoma / Q000032:analysis* / Q000473:pathology; D002460:Cell Line; D002465:Cell Movement; D002630:Chemotactic Factors / Q000302:isolation & purification* / Q000502:physiology; D002634:Chemotaxis, Leukocyte; D006801:Humans; D016209:Interleukin-8; D008175:Lung Neoplasms / Q000032:analysis* / Q000473:pathology; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000302:isolation & purification*; D009504:Neutrophils / Q000502:physiology*; D012689:Sequence Homology, Nucleic Acid
D000596:Amino Acids; D002630:Chemotactic Factors; D016209:Interleukin-8; D009363:Neoplasm Proteins