The antibody sc-33040-R fails to specifically recognize phosphorylation of ErbB4 on tyrosine1056

Gallo, Richard M.; Riese, David J.

doi:10.1080/08977190701804008

Cited by 4 publications

(4 citation statements)

References 36 publications

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“…We postulated that the failure of NRG2α/K45F and NRG2β/Q43L to stimulate ErbB4 coupling to IL-3-independent proliferation is a result of their failure to stimulate phosphorylation of critical ErbB4 tyrosine residues. However, the availability of ErbB4 anti-phosphotyrosine antibodies is limited [28], and MS approaches to mapping sites of ErbB4 tyrosine phosphorylation have been limited to studies of ligand-independent ErbB4 tyrosine phosphorylation [29–31]. Instead, we have postulated that the maximal level of ErbB4 tyrosine phosphorylation stimulated by agonistic and antagonistic ErbB4 ligands is different, indicating that agonistic ErbB4 ligands are stimulating phosphorylation of more tyrosine residues (per ErbB4 molecule) than are antagonistic ligands.…”

Section: Resultsmentioning

confidence: 99%

The Q43L mutant of neuregulin 2β is a pan-ErbB receptor antagonist

Wilson

Mill

Gallo

et al. 2012

Biochemical Journal

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The ErbB4 receptor tyrosine kinase possesses both tumour suppressor and oncogenic activities. Thus pharmacological agents are needed to help elucidate ErbB4 functions. However, limitations of existing ErbB4 agonists and antagonists have led us to seek novel ErbB4 antagonists. The Q43L mutant of the ErbB4 agonist NRG2β (neuregulin 2β) stimulates ErbB4 tyrosine phosphorylation, yet fails to stimulate ErbB4 coupling to cell proliferation. Thus in the present paper we hypothesize that NRG2β/Q43L may be an ErbB4 antagonist. NRG2β/Q43L competitively antagonizes agonist stimulation of ErbB4 coupling to cell proliferation. NRG2β/Q43L stimulates less ErbB4 tyrosine phosphorylation than does NRG2β. In addition, NRG2β stimulation of cell proliferation requires PI3K (phosphoinositide 3-kinase) activity and NRG2β stimulates greater Akt phosphorylation than does NRG2β/Q43L. Moreover, EGFR [EGF (epidermal growth factor) receptor] kinase activity (but not that of ErbB4) is critical for coupling ErbB4 to proliferation. Experiments utilizing ErbB4 splicing isoforms and mutants suggest that NRG2β and NRG2β/Q43L may differentially stimulate ErbB4 coupling to the transcriptional co-regulator YAP (Yes-associated protein). Finally, NRG2β/Q43L competitively antagonizes agonist stimulation of EGFR and ErbB2/ErbB3, indicating that NRG2β/Q43L is a pan-ErbB antagonist. Thus we postulate that NRG2β/Q43L and other antagonistic ligands stimulate ErbB tyrosine phosphorylation on a set of residues distinct from that stimulated by agonists, thus suggesting a novel mechanism of ErbB receptor regulation. Moreover, NRG2β/Q43L and related ligand-based antagonists establish a paradigm for the discovery of anti-ErbB therapeutics.

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Section: Resultsmentioning

confidence: 99%

The Q43L mutant of neuregulin 2β is a pan-ErbB receptor antagonist

Wilson

Mill

Gallo

et al. 2012

Biochemical Journal

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“…Sample buffer lysates normalized for protein concentration were analyzed by electrophoresis in 4–12% NuPAGE SDS-polyacrylamide midigels (Life Technologies Corporation). For immunoblotting, PVDF membranes were blocked with 2% BSA in 10 mM Tris-HCl,50 mM NaCl, 0.1% Tween 20, pH 7.4 (TBST) and incubated with anti- phospho-ERBB4 (Tyr 1056) (22), phospho-ERBB4 Tyr 1284 (Cell Signaling Technology #4757), ERBB4 (sc-283), GAPDH (Santa Cruz Biotechnology, CA), phospho-MAPK (Thr202/Tyr204), or phospho-AKT (Ser473; Cell Signaling Technology, MA) diluted 1:5000 to 1:20000 in TBST/2% BSA for 2 h. Membranes were washed five times with TBST, incubated with horseradish peroxide-conjugated secondary antibodies in TBST/2% BSA for 1 h, rinsed with TBST, and detected by chemiluminescence (SuperSignal West Pico Chemiluminescent Substrate; Pierce, Rockford, IL).…”

Section: Methodsmentioning

confidence: 99%

Convergent and Divergent Cellular Responses by ErbB4 Isoforms in Mammary Epithelial Cells

Wali

Haskins

Gilmore-Hebert

et al. 2014

Molecular Cancer Research

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Associations of ErbB4 (ERBB4/HER4), the fourth member of the EGFR family, with cancer are variable, possibly as a result of structural diversity of this receptor. There are multiple structural isoforms of ERBB4 arising by alternative mRNA splicing, and a subset undergo proteolysis that releases membrane-anchored and soluble isoforms that associate with transcription factors and co-regulators to modulate transcription. In order to compare the differential and common signaling activities of full-length and soluble intracellular isoforms of ERBB4, four JM-a isoforms (full-length and soluble intracellular domain CYT-1 and CYT-2) were expressed in isogenic MCF10A cells and their biological activities were analyzed. Both full-length and intracellular domain CYT-2 promoted cell proliferation and invasion, and CYT-1 suppressed cell growth. Transcriptional profiling revealed several new and underexplored ERBB4 regulated transcripts including: proteases/protease inhibitors (MMP3, SERPINE2), YAP/Hippo pathway (CTGF, CYR61, SPARC), mevalonate/cholesterol pathway (HMGCR, HMGCS1, LDLR, DHCR7), and cytokines (IL8, CCL20, CXCL1). Many of these transcripts were subsequently validated in a luminal, breast cancer cell line that normally express ERBB4. Furthermore, ChiP-seq experiments identified ADAP1, APOE, SPARC, STMN1, and MXD1 as novel molecular targets of ERBB4. These findings clarify the diverse biological activities of ERBB4 isoforms, and reveal new and divergent functions.

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“…Additional splicing results in alterations to the cytoplasmic domain (towards the C-terminal tail), producing the CYT-1 and CYT-2 variants by the inclusion or exclusion of exon 26 ( 9 ). This results in the respective presence or absence of a 16-amino acid sequence (Ser1046 through Gly1061) ( 11 – 13 ).…”

Section: Erbb4 Domain Structurementioning

confidence: 99%

“…Whilst ErbB4 is activated by a specific subset of ligands, once ligand-bound, it can homodimerize with other ErbB4 receptor molecules or heterodimerize with other ErbB family members. This complicates the study of ErbB4-induced signalling events ( 13 ). Compounding this, the lack of ErbB4-specific reagents can make it challenging to study its biological role.…”

Section: The Lack Of Isoform-specific and Cleavage-specific Reagentsmentioning

confidence: 99%

ErbB4 in the brain: Focus on high grade glioma

Pitcher

Alexander²,

Miranda³

et al. 2022

Front. Oncol.

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The epidermal growth factor receptor (EGFR) family of receptor tyrosine kinases (RTKs) consists of EGFR, ErbB2, ErbB3, and ErbB4. These receptors play key roles in cell proliferation, angiogenesis, cell migration, and in some cases, tumor promotion. ErbB4 is a unique member of the EGFR family, implicated not only in pro-tumorigenic mechanisms, such as cell proliferation and migration, but also in anti-tumorigenic activities, including cell differentiation and apoptosis. ErbB4 is differentially expressed in a wide variety of tissues, and interestingly, as different isoforms that result in vastly different signalling outcomes. Most studies have either ignored the presence of these isoforms or used overexpression models that may mask the true function of ErbB4. ErbB4 is widely expressed throughout the body with significant expression in skeletal tissue, mammary glands, heart, and brain. Knockout models have demonstrated embryonic lethality due to disrupted heart and brain development. Despite high expression in the brain and a critical role in brain development, remarkably little is known about the potential signalling activity of ErbB4 in brain cancer.This review focuses on the unique biology of ErbB4 in the brain, and in particular, highlights brain cancer research findings. We end the review with a focus on high grade gliomas, primarily glioblastoma, a disease that has been shown to involve EGFR and its mutant forms. The role of the different ErbB4 isotypes in high grade gliomas is still unclear and future research will hopefully shed some light on this question.

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The antibody sc-33040-R fails to specifically recognize phosphorylation of ErbB4 on tyrosine1056

Cited by 4 publications

References 36 publications

The Q43L mutant of neuregulin 2β is a pan-ErbB receptor antagonist

The Q43L mutant of neuregulin 2β is a pan-ErbB receptor antagonist

Convergent and Divergent Cellular Responses by ErbB4 Isoforms in Mammary Epithelial Cells

ErbB4 in the brain: Focus on high grade glioma

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