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Protein phosphorylation of neutrophils from normal children and patients with chronic granulomatous disease

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Abstract

The phosphorylation and dephosphorylation of proteins in neutrophils from normal children and patients with chronic granulomatous disease (CGD) were studied with two-dimensional gel electrophoresis and autoradiography, followed by densitometric scanning. In normal neutrophils the radio-activities of 11 spots among approximately 50 radioactive spots were changed by stimulation with phorbol 12-myristate 13-acetate (PMA) and 6 of the 11 spots were also changed by stimulation with N-formyl-methionyl-leucyl-phenylalanine (FMLP) and NaF. The phosphorylation of only two spots (Mr=48000 and 62000) was inhibited by 2-deoxyglucose and N-(6-aminohexyl)-5-chloro-1-naphthalene sulphonamide (W-7), which inhibits superoxide production, while it was not affected by dibutyryl cAMP, KCN and ethyleneglycol-bis-(β-aminoethylether)-N,N′-tetraacetic acid (EGTA), which do not affect superoxide production. The observation indicates that the Mr=48000 and 62000 proteins may be involved in the activation process of superoxide production. When the neutrophils of four male and two female CGD patients were examined, the changes in 11 spots on stimulation were similar to those of normal children, indicating that the (de)phosphorylation of the proteins which seems to be involved in the activation process is not affected in CGD neutrophils.

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Abbreviations

CGD:

chronic granulomatous disease

PMA:

phorbol 12-myristate 13-acetate

FMLP:

N-formyl-methionylleucyl-phenylalanine

W-7:

N-(6-aminohexyl)-5-chloro-1-naphthalane sulphonamide

EGTA:

ethyleneglycol-bis-(β-aminoethylether)-N,N′-tetraacetic acid

dBcAMP:

dibutyryl cyclic AMP

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Authors and Affiliations

  1. Department of Pediatrics, Kyushu University School of Medicine, Maidashi 3-1-1, Higashi-ku, 812, Fukuoka, Japan

    E. Ishii & K. Ueda

  2. Biochemistry, Kyushu University School of Medicine, Fukuoka, Japan

    K. Irita, I. Fujita & K. Takeshige

  3. Department of Pediatrics, Hiroshima University School of Medicine, Hiroshima, Japan

    M. Kobayashi & T. Usui

Authors
  1. E. Ishii
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  2. K. Irita
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  3. I. Fujita
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  4. K. Takeshige
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  5. M. Kobayashi
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  6. T. Usui
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  7. K. Ueda
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Ishii, E., Irita, K., Fujita, I. et al. Protein phosphorylation of neutrophils from normal children and patients with chronic granulomatous disease. Eur J Pediatr 145, 22–26 (1986). https://doi.org/10.1007/BF00441847

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  • DOI: https://doi.org/10.1007/BF00441847

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