Activation-dependent clustering of the erbB2 receptor tyrosine kinase detected by scanning near-field optical microscopy

Péter Nagy, A. Jenei, Achim K. Kirsch, J. Szöllősi, S. Damjanovich, Thomas M. Jovin

Research output: Contribution to journalArticle

110 Citations (Scopus)

Abstract

ErbB2 (HER2, Neu), a member of the epidermal growth factor (EGF) receptor tyrosine kinase family, is often overexpressed in breast cancer and other malignancies. ErbB2 homodimerizes but also presents as a common auxiliary subunit of the EGF acid heregulin receptors (erbB1 or EGFR; and erbB3-4, respectively), with which it heteroassociates. ErbB2 is generally regarded as an orphan (ligand-less) receptor with a very potent kinase domain activated either via its associated partners or constitutively as a consequence of discrete mutations. It follows that the extent and regulation of its cell surface interactions are of central importance. We have studied the large-scale association pattern of erbB2 in quiescent and activated cells labeled with fluorescent anti-erbB2 monoclonal antibodies using scanning near-field optical microscopy (SNOM). ErbB2 was found to be concentrated in irregular membrane patches with a mean diameter of approx. 0.5 μm in nonactivated SKBR3 and MDA453 human breast tumor cells. The average number of erbB2 proteins in a single cluster on nonactivated SKBR3 cells was about 103. Activation of SKBR3 cells with EGF, heregulin as well as a partially agonistic anti-erbB2 monoclonal antibody led to an increase in the mean cluster diameter to 0.6-0.9 μm, irrespective of the ligand. The EGF-induced increase in the erbB2 cluster size was inhibited by the EGFR-specific tyrosine kinase inhibitor PD153035. The average size of erbB2 clusters on the erbB2-transfected line of CHO cells (CB2) was similar to that of activated SKBR3 cells, a finding correlated with the increased base-line tyrosine phosphorylation of erbB2 in cells expressing only erbB2. We conclude that an increase in cluster size may constitute a general phenomenon in the activation of erbB2.

Original languageEnglish
Pages (from-to)1733-1741
Number of pages9
JournalJournal of Cell Science
Volume112
Issue number11
Publication statusPublished - 1999

Fingerprint

Receptor Protein-Tyrosine Kinases
Cluster Analysis
Microscopy
Epidermal Growth Factor
Neuregulin-1
Protein-Tyrosine Kinases
Monoclonal Antibodies
Breast Neoplasms
Ligands
Orphaned Children
CHO Cells
Epidermal Growth Factor Receptor
Cell Communication
Tyrosine
Phosphotransferases
Phosphorylation
Mutation
Acids
Membranes
Neoplasms

Keywords

  • EGF receptor
  • ErbB2
  • Homassociation
  • NSOM
  • SNOM

ASJC Scopus subject areas

  • Cell Biology

Cite this

Activation-dependent clustering of the erbB2 receptor tyrosine kinase detected by scanning near-field optical microscopy. / Nagy, Péter; Jenei, A.; Kirsch, Achim K.; Szöllősi, J.; Damjanovich, S.; Jovin, Thomas M.

In: Journal of Cell Science, Vol. 112, No. 11, 1999, p. 1733-1741.

Research output: Contribution to journalArticle

@article{6b023f9e2dbf42699e96532509ede646,
title = "Activation-dependent clustering of the erbB2 receptor tyrosine kinase detected by scanning near-field optical microscopy",
abstract = "ErbB2 (HER2, Neu), a member of the epidermal growth factor (EGF) receptor tyrosine kinase family, is often overexpressed in breast cancer and other malignancies. ErbB2 homodimerizes but also presents as a common auxiliary subunit of the EGF acid heregulin receptors (erbB1 or EGFR; and erbB3-4, respectively), with which it heteroassociates. ErbB2 is generally regarded as an orphan (ligand-less) receptor with a very potent kinase domain activated either via its associated partners or constitutively as a consequence of discrete mutations. It follows that the extent and regulation of its cell surface interactions are of central importance. We have studied the large-scale association pattern of erbB2 in quiescent and activated cells labeled with fluorescent anti-erbB2 monoclonal antibodies using scanning near-field optical microscopy (SNOM). ErbB2 was found to be concentrated in irregular membrane patches with a mean diameter of approx. 0.5 μm in nonactivated SKBR3 and MDA453 human breast tumor cells. The average number of erbB2 proteins in a single cluster on nonactivated SKBR3 cells was about 103. Activation of SKBR3 cells with EGF, heregulin as well as a partially agonistic anti-erbB2 monoclonal antibody led to an increase in the mean cluster diameter to 0.6-0.9 μm, irrespective of the ligand. The EGF-induced increase in the erbB2 cluster size was inhibited by the EGFR-specific tyrosine kinase inhibitor PD153035. The average size of erbB2 clusters on the erbB2-transfected line of CHO cells (CB2) was similar to that of activated SKBR3 cells, a finding correlated with the increased base-line tyrosine phosphorylation of erbB2 in cells expressing only erbB2. We conclude that an increase in cluster size may constitute a general phenomenon in the activation of erbB2.",
keywords = "EGF receptor, ErbB2, Homassociation, NSOM, SNOM",
author = "P{\'e}ter Nagy and A. Jenei and Kirsch, {Achim K.} and J. Sz{\"o}llősi and S. Damjanovich and Jovin, {Thomas M.}",
year = "1999",
language = "English",
volume = "112",
pages = "1733--1741",
journal = "Journal of Cell Science",
issn = "0021-9533",
publisher = "Company of Biologists Ltd",
number = "11",

}

TY - JOUR

T1 - Activation-dependent clustering of the erbB2 receptor tyrosine kinase detected by scanning near-field optical microscopy

AU - Nagy, Péter

AU - Jenei, A.

AU - Kirsch, Achim K.

AU - Szöllősi, J.

AU - Damjanovich, S.

AU - Jovin, Thomas M.

PY - 1999

Y1 - 1999

N2 - ErbB2 (HER2, Neu), a member of the epidermal growth factor (EGF) receptor tyrosine kinase family, is often overexpressed in breast cancer and other malignancies. ErbB2 homodimerizes but also presents as a common auxiliary subunit of the EGF acid heregulin receptors (erbB1 or EGFR; and erbB3-4, respectively), with which it heteroassociates. ErbB2 is generally regarded as an orphan (ligand-less) receptor with a very potent kinase domain activated either via its associated partners or constitutively as a consequence of discrete mutations. It follows that the extent and regulation of its cell surface interactions are of central importance. We have studied the large-scale association pattern of erbB2 in quiescent and activated cells labeled with fluorescent anti-erbB2 monoclonal antibodies using scanning near-field optical microscopy (SNOM). ErbB2 was found to be concentrated in irregular membrane patches with a mean diameter of approx. 0.5 μm in nonactivated SKBR3 and MDA453 human breast tumor cells. The average number of erbB2 proteins in a single cluster on nonactivated SKBR3 cells was about 103. Activation of SKBR3 cells with EGF, heregulin as well as a partially agonistic anti-erbB2 monoclonal antibody led to an increase in the mean cluster diameter to 0.6-0.9 μm, irrespective of the ligand. The EGF-induced increase in the erbB2 cluster size was inhibited by the EGFR-specific tyrosine kinase inhibitor PD153035. The average size of erbB2 clusters on the erbB2-transfected line of CHO cells (CB2) was similar to that of activated SKBR3 cells, a finding correlated with the increased base-line tyrosine phosphorylation of erbB2 in cells expressing only erbB2. We conclude that an increase in cluster size may constitute a general phenomenon in the activation of erbB2.

AB - ErbB2 (HER2, Neu), a member of the epidermal growth factor (EGF) receptor tyrosine kinase family, is often overexpressed in breast cancer and other malignancies. ErbB2 homodimerizes but also presents as a common auxiliary subunit of the EGF acid heregulin receptors (erbB1 or EGFR; and erbB3-4, respectively), with which it heteroassociates. ErbB2 is generally regarded as an orphan (ligand-less) receptor with a very potent kinase domain activated either via its associated partners or constitutively as a consequence of discrete mutations. It follows that the extent and regulation of its cell surface interactions are of central importance. We have studied the large-scale association pattern of erbB2 in quiescent and activated cells labeled with fluorescent anti-erbB2 monoclonal antibodies using scanning near-field optical microscopy (SNOM). ErbB2 was found to be concentrated in irregular membrane patches with a mean diameter of approx. 0.5 μm in nonactivated SKBR3 and MDA453 human breast tumor cells. The average number of erbB2 proteins in a single cluster on nonactivated SKBR3 cells was about 103. Activation of SKBR3 cells with EGF, heregulin as well as a partially agonistic anti-erbB2 monoclonal antibody led to an increase in the mean cluster diameter to 0.6-0.9 μm, irrespective of the ligand. The EGF-induced increase in the erbB2 cluster size was inhibited by the EGFR-specific tyrosine kinase inhibitor PD153035. The average size of erbB2 clusters on the erbB2-transfected line of CHO cells (CB2) was similar to that of activated SKBR3 cells, a finding correlated with the increased base-line tyrosine phosphorylation of erbB2 in cells expressing only erbB2. We conclude that an increase in cluster size may constitute a general phenomenon in the activation of erbB2.

KW - EGF receptor

KW - ErbB2

KW - Homassociation

KW - NSOM

KW - SNOM

UR - http://www.scopus.com/inward/record.url?scp=0033014064&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033014064&partnerID=8YFLogxK

M3 - Article

C2 - 10318765

AN - SCOPUS:0033014064

VL - 112

SP - 1733

EP - 1741

JO - Journal of Cell Science

JF - Journal of Cell Science

SN - 0021-9533

IS - 11

ER -