Characterization of a 64-kd protein phosphorylated during chemotactic activation with IL-8 and fMLP of human polymorphonuclear leukocytes. II. Purification and amino acid analysis of phosphorylated 64-kd protein.
Lung giant cell carcinoma-derived chemotactic protein (LUCT)/IL-8 and fMet-Leu-Phe stimulate phosphorylation of a 64-kd protein (p64) in 32P-labeled human polymorphonuclear leukocytes (PMNs). The p64 was purified from cytosol of human PMNs (1.8 x 10(9) cells) by DEAE-Sepharose CL6B column chromatography, hydroxyapatite HPLC, and reverse-phase HPLC. By hydroxyapatite HPLC, p64s were separated and produced two peaks containing both nonphosphorylated and phosphorylated p64. Amino acid composition of each p64 was determined. Each p64 was directly subjected to amino acid sequencing, but the amino acid residue in the amino-terminal of the proteins could not be detected. From the results of amino acid composition and other characters of p64, it is suggested that p64 is identical to l-plastin, which is a leukocyte-specific protein.
Shibata
M
M
Department of Bioactive Molecules, National Institute of Health, Tokyo, Japan.
Yamakawa
Y
Y
Ohoka
T
T
Mizuno
S
S
Suzuki
K
K
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis*; D002630:Chemotactic Factors / Q000494:pharmacology*; D003600:Cytosol / Q000737:chemistry; D006801:Humans; D016209:Interleukin-8 / Q000494:pharmacology*; D008562:Membrane Glycoproteins; D008840:Microfilament Proteins; D008969:Molecular Sequence Data; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology*; D009504:Neutrophils / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D010750:Phosphoproteins / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation / Q000187:drug effects; D013347:Subcellular Fractions / Q000737:chemistry
D000596:Amino Acids; D002630:Chemotactic Factors; D016209:Interleukin-8; D008562:Membrane Glycoproteins; D008840:Microfilament Proteins; D010750:Phosphoproteins; D010761:Phosphorus Radioisotopes; C026742:plastin; D009240:N-Formylmethionine Leucyl-Phenylalanine