Heregulin reverses the oligomerization of HER3

Ralf Landgraf, David Eisenberg

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

We analyzed the propensity of the HER3 receptor and its extracellular domain (ECD) to undergo ligand-independent self-association. The HER3-ECD, purified from Drosophila S2 cells, binds the EGF-like domain of heregulin (hrg) with a K(d) of 1.9 nM as measured by surface plasmon resonance (SPR) studies. In a gel shift assay, the HER3-ECD self-associates into a uniform, slowly migrating species in a concentration-dependent manner, starting at concentrations of < 10 nM. In contrast to the HER3-ECD, the ECD from the related HER2 receptor does not oligomerize under the same conditions. The direct interaction of HER3-ECDs was also demonstrated by pull-down assays and SPR measurements under physiological salt conditions. This self-association of the HER3-ECD was reversed by the addition of hrg but not by EGF. The apparent equilibrium dissociation constant for the HER3-ECD self-association is 15 nM, based on SPR measurements. In this analysis, hrg blocks HER3-ECD self-association, and the addition of hrg during the dissociation phase resulted in an accelerated off rate. This finding suggests that hrg can bind to and disrupt preexisting HER3-ECD oligomers. Full-length HER3 likewise exhibited self-association. Under conditions where co-immunoprecipitation and cross-linking of HER2 and HER3 were stimulated by hrg, HER3 self-association and cross-linking were disrupted by hrg. The implication is that the self- association of HER3-ECD favors the formation of catalytically inactive complexes of the HER3 receptor. Binding of hrg releases HER3 which may then form signaling-competent HER3-HER2 heterodimers.

Original languageEnglish
Pages (from-to)8503-8511
Number of pages9
JournalBiochemistry
Volume39
Issue number29
DOIs
StatePublished - Jul 25 2000
Externally publishedYes

Fingerprint

Neuregulin-1
Oligomerization
Association reactions
Surface Plasmon Resonance
Surface plasmon resonance
Epidermal Growth Factor
Assays
Immunoprecipitation
Oligomers
Drosophila
Salts
Gels
Ligands

ASJC Scopus subject areas

  • Biochemistry

Cite this

Heregulin reverses the oligomerization of HER3. / Landgraf, Ralf; Eisenberg, David.

In: Biochemistry, Vol. 39, No. 29, 25.07.2000, p. 8503-8511.

Research output: Contribution to journalArticle

Landgraf, Ralf ; Eisenberg, David. / Heregulin reverses the oligomerization of HER3. In: Biochemistry. 2000 ; Vol. 39, No. 29. pp. 8503-8511.
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