Novel association of the Src family kinases, Hck and c-Fgr, with CCR3 receptor stimulation: A possible mechanism for eotaxin-induced human eosinophil chemotaxis

Amr El-Shazly, Naoto Yamaguchi, Keisuke Masuyama, Toshio Suda, Takeru Ishikawa

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

The chemokine eotaxin is a potent and relatively eosinophil-specific chemoattractant implicated in the cell migration to inflammatory sites in allergic diseases. Eotaxin exerts its activity solely through the CCR3 receptor, but the signaling pathways are poorly defined. In this study, we show that eotaxin induces an increase in tyrosine phosphorylation of multiple cellular proteins in normal human eosinophils. Eotaxin-dependent tyrosine phosphorylation was detected 1 min after stimulation and increased for at least 15 min with kinetics similar to those of eotaxin-induced cell shape changes. Herbimycin A, a tyrosine kinase inhibitor, blocked both eotaxin-induced tyrosine phosphorylation and cell shape changes as well as chemotaxis. Immunofluorescence microscopy analyses showed that eotaxin-induced cell shape changes were accompanied by redistribution of tyrosine-phosphorylated proteins and F-actin reorganization that were sensitive to herbimycin A. Coimmunoprecipitation studies revealed that binding of eotaxin to CCR3 greatly enhanced association of the Src family kinases, Hck and c-Fgr, with CCR3 after internalization of CCR3. These results may indicate that recruitment of Hck and c-Fgr to CCR3 in a compartment triggers tyrosine phosphorylation, leading to rapid cell shape changes required for cell migration.

Original languageEnglish
Pages (from-to)163-170
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume264
Issue number1
DOIs
Publication statusPublished - 1999 Oct 14

Fingerprint

CCR3 Receptors
src-Family Kinases
Chemotaxis
Eosinophils
Phosphorylation
Cell Shape
Tyrosine
Association reactions
Cell Movement
Chemotactic Factors
Fluorescence Microscopy
Chemokines
Protein-Tyrosine Kinases
Actins
Microscopic examination
Proteins
Kinetics

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Novel association of the Src family kinases, Hck and c-Fgr, with CCR3 receptor stimulation : A possible mechanism for eotaxin-induced human eosinophil chemotaxis. / El-Shazly, Amr; Yamaguchi, Naoto; Masuyama, Keisuke; Suda, Toshio; Ishikawa, Takeru.

In: Biochemical and Biophysical Research Communications, Vol. 264, No. 1, 14.10.1999, p. 163-170.

Research output: Contribution to journalArticle

@article{85307ddbea5b48a59b60fac32a491e3b,
title = "Novel association of the Src family kinases, Hck and c-Fgr, with CCR3 receptor stimulation: A possible mechanism for eotaxin-induced human eosinophil chemotaxis",
abstract = "The chemokine eotaxin is a potent and relatively eosinophil-specific chemoattractant implicated in the cell migration to inflammatory sites in allergic diseases. Eotaxin exerts its activity solely through the CCR3 receptor, but the signaling pathways are poorly defined. In this study, we show that eotaxin induces an increase in tyrosine phosphorylation of multiple cellular proteins in normal human eosinophils. Eotaxin-dependent tyrosine phosphorylation was detected 1 min after stimulation and increased for at least 15 min with kinetics similar to those of eotaxin-induced cell shape changes. Herbimycin A, a tyrosine kinase inhibitor, blocked both eotaxin-induced tyrosine phosphorylation and cell shape changes as well as chemotaxis. Immunofluorescence microscopy analyses showed that eotaxin-induced cell shape changes were accompanied by redistribution of tyrosine-phosphorylated proteins and F-actin reorganization that were sensitive to herbimycin A. Coimmunoprecipitation studies revealed that binding of eotaxin to CCR3 greatly enhanced association of the Src family kinases, Hck and c-Fgr, with CCR3 after internalization of CCR3. These results may indicate that recruitment of Hck and c-Fgr to CCR3 in a compartment triggers tyrosine phosphorylation, leading to rapid cell shape changes required for cell migration.",
author = "Amr El-Shazly and Naoto Yamaguchi and Keisuke Masuyama and Toshio Suda and Takeru Ishikawa",
year = "1999",
month = "10",
day = "14",
doi = "10.1006/bbrc.1999.1379",
language = "English",
volume = "264",
pages = "163--170",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Novel association of the Src family kinases, Hck and c-Fgr, with CCR3 receptor stimulation

T2 - A possible mechanism for eotaxin-induced human eosinophil chemotaxis

AU - El-Shazly, Amr

AU - Yamaguchi, Naoto

AU - Masuyama, Keisuke

AU - Suda, Toshio

AU - Ishikawa, Takeru

PY - 1999/10/14

Y1 - 1999/10/14

N2 - The chemokine eotaxin is a potent and relatively eosinophil-specific chemoattractant implicated in the cell migration to inflammatory sites in allergic diseases. Eotaxin exerts its activity solely through the CCR3 receptor, but the signaling pathways are poorly defined. In this study, we show that eotaxin induces an increase in tyrosine phosphorylation of multiple cellular proteins in normal human eosinophils. Eotaxin-dependent tyrosine phosphorylation was detected 1 min after stimulation and increased for at least 15 min with kinetics similar to those of eotaxin-induced cell shape changes. Herbimycin A, a tyrosine kinase inhibitor, blocked both eotaxin-induced tyrosine phosphorylation and cell shape changes as well as chemotaxis. Immunofluorescence microscopy analyses showed that eotaxin-induced cell shape changes were accompanied by redistribution of tyrosine-phosphorylated proteins and F-actin reorganization that were sensitive to herbimycin A. Coimmunoprecipitation studies revealed that binding of eotaxin to CCR3 greatly enhanced association of the Src family kinases, Hck and c-Fgr, with CCR3 after internalization of CCR3. These results may indicate that recruitment of Hck and c-Fgr to CCR3 in a compartment triggers tyrosine phosphorylation, leading to rapid cell shape changes required for cell migration.

AB - The chemokine eotaxin is a potent and relatively eosinophil-specific chemoattractant implicated in the cell migration to inflammatory sites in allergic diseases. Eotaxin exerts its activity solely through the CCR3 receptor, but the signaling pathways are poorly defined. In this study, we show that eotaxin induces an increase in tyrosine phosphorylation of multiple cellular proteins in normal human eosinophils. Eotaxin-dependent tyrosine phosphorylation was detected 1 min after stimulation and increased for at least 15 min with kinetics similar to those of eotaxin-induced cell shape changes. Herbimycin A, a tyrosine kinase inhibitor, blocked both eotaxin-induced tyrosine phosphorylation and cell shape changes as well as chemotaxis. Immunofluorescence microscopy analyses showed that eotaxin-induced cell shape changes were accompanied by redistribution of tyrosine-phosphorylated proteins and F-actin reorganization that were sensitive to herbimycin A. Coimmunoprecipitation studies revealed that binding of eotaxin to CCR3 greatly enhanced association of the Src family kinases, Hck and c-Fgr, with CCR3 after internalization of CCR3. These results may indicate that recruitment of Hck and c-Fgr to CCR3 in a compartment triggers tyrosine phosphorylation, leading to rapid cell shape changes required for cell migration.

UR - http://www.scopus.com/inward/record.url?scp=0033554561&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033554561&partnerID=8YFLogxK

U2 - 10.1006/bbrc.1999.1379

DO - 10.1006/bbrc.1999.1379

M3 - Article

C2 - 10527858

AN - SCOPUS:0033554561

VL - 264

SP - 163

EP - 170

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 1

ER -