Detection of ERK activation by a novel monoclonal antibody

Yung, Yuval; Dolginov, Yakov; Yao, Zhong; Rubinfeld, Hadara; Michael, Dan; Hanoch, Tamar; Roubini, Eli; Lando, Zeev; Zharhary, Dorit; Seger, Rony

doi:10.1016/s0014-5793(97)00442-0

Cited by 130 publications

(99 citation statements)

References 25 publications

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“…Erk2 is a serine/threonine kinase, which is activated by sequential phosphorylation on tyrosine and threonine residues (Yung et al, 1997). Although data presented here demonstrate a significant effect of TSP-1 on tyrosine phosphorylation of Erk2, this does not necessarily reflect an increase in the activity of the enzyme.…”

Section: Analysis Of Extracellular-regulated Kinase Activitycontrasting

confidence: 68%

Thrombospondin-1 differentially induces chemotaxis and DNA synthesis of human venous smooth muscle cells at the receptor-binding level

Lymn

Patel

Clunn

et al. 2002

Journal of Cell Science

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Thrombospondin-1 is a large matricellular protein that acts as a pleiotropic growth factor for human vascular smooth muscle cells, and may play a role in the progression of vascular disease. Although we have previously demonstrated the dependence of both thrombospondin-1-stimulated cell chemotaxis and proliferation on tyrosine kinases, the receptor mechanisms involved remain obscure. This investigation aims to determine the nature of the receptor(s) involved in the cellular responses to thrombospondin-1. Cellular signals were identified by western blotting following cell stimulation, while cellular responses were assessed by measuring DNA synthesis and chemotaxis. These data demonstrate that thrombospondin-1-induced cell chemotaxis can be inhibited by a peptide containing the Arg-Gly-Asp motif, a function-blocking αvβ3 antibody, a function-blocking integrin-associated protein (IAP) antibody and pertussis toxin, while thrombospondin-1-stimulated DNA synthesis is inhibited by a function-blocking α3β1 antibody. Similarly the Arg-Gly-Asp-containing peptide inhibits tyrosine phosphorylation of focal adhesion kinase and the p85 regulatory subunit of phosphatidylinositol 3-kinase, but does not significantly affect tyrosine phosphorylation, or activation, of extracellular-regulated kinase. These data suggest that soluble thrombospondin-1 interacts with human vascular smooth muscle cells via two independent and separable receptor-binding sites, to differentially stimulate cell chemotaxis and DNA synthesis.

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Section: Analysis Of Extracellular-regulated Kinase Activitycontrasting

confidence: 68%

Thrombospondin-1 differentially induces chemotaxis and DNA synthesis of human venous smooth muscle cells at the receptor-binding level

Lymn

Patel

Clunn

et al. 2002

Journal of Cell Science

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“…The PY20 antibody was purchased from Santa Cruz Biotechnology. A mAb to the active form of the MAP kinase (Yung et al, 1997) was a gift from R Seger (Weizmann Institute).…”

Section: Methodsmentioning

confidence: 99%

Neuregulin-4: a novel growth factor that acts through the ErbB-4 receptor tyrosine kinase

et al. 1999

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The ErbB/HER family of receptor tyrosine kinases consists of four receptors that bind a large number of growth factor ligands sharing an epidermal growth factor-(EGF)-like motif. Whereas ErbB-1 binds seven di erent ligands whose prototype is EGF, the three families of neuregulins (NRGs) activate ErbB-3 and/or ErbB-4. Here we characterize a fourth neuregulin, NRG-4, that acts through ErbB-4. The predicted pro-NRG-4 is a transmembrane protein carrying a unique EGF-like motif and a short cytoplasmic domain. A synthetic peptide encompassing the full-length EGF-like domain can induce growth of interleukin-dependent cells ectopically expressing ErbB-4, but not cells expressing the other three ErbB proteins or their combinations. Consistent with speci®city to ErbB-4, NRG-4 can displace an ErbB-4-bound NRG-1 and can activate signaling downstream of this receptor. Expression of NRG-4 mRNA was detected in the adult pancreas and weakly in muscle; other tissues displayed no detectable NRG-4 mRNA. The primary structure and the pattern of expression of NRG-4, together with the strict speci®city of this growth factor to ErbB-4, suggest a physiological role distinct from that of the known ErbB ligands.

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“…The biological differences we observed between epiregulin, EGF, and NDF in subsets of 32D cells suggested that these ligands may differ in signaling potency, and especially in their ability to recruit the MAPK pathway. To analyze receptor phosphorylation and MAPK activation we probed blots of whole extracts, prepared from ligandstimulated cells, with antibodies to phosphotyrosine, or with a murine mAb that specifically recognizes the active, doubly phosphorylated form of the ERK1 and ERK2 MAPKs (26). Surprisingly, the more mitogenic ligand of ErbB-1, epiregulin, exhibited weaker, but not less sustained, tyrosine phosphorylation of proteins at the 180-kDa range corresponding to ErbB-1 in D1 cells (Fig.…”

Section: Epiregulin Is a Relatively Potent Stimulator Of Erbb-1 But mentioning

confidence: 99%

Epiregulin Is a Potent Pan-ErbB Ligand That Preferentially Activates Heterodimeric Receptor Complexes

Shelly¹,

Pinkas‐Kramarski²,

Guarino³

et al. 1998

Journal of Biological Chemistry

147

121

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The ErbB signaling network consists of four transmembrane receptor tyrosine kinases and more than a dozen ligands sharing an epidermal growth factor (EGF) motif. The multiplicity of ErbB-specific ligands is incompletely understood in terms of signal specificity because all ErbB molecules signal through partially overlapping pathways. Here we addressed the action of epiregulin, a recently isolated ligand of ErbB-1. By employing a set of factor-dependent cell lines engineered to express individual ErbBs or their combinations, we found that epiregulin is the broadest specificity EGF-like ligand so far characterized: not only does it stimulate homodimers of both ErbB-1 and ErbB-4, it also activates all possible heterodimeric ErbB complexes. Consistent with its relaxed selectivity, epiregulin binds the various receptor combinations with an affinity that is approximately 100-fold lower than the affinity of ligands with more stringent selectivity, including EGF. Nevertheless, epiregulin's action upon most receptor combinations transmits a more potent mitogenic signal than does EGF. This remarkable discrepancy between binding affinity and bioactivity is permitted by a mechanism that prevents receptor down-regulation, and results in a weak, but prolonged, state of receptor activation.Various biological processes are controlled by intercellular interactions that are mediated by polypeptide growth factors. Examples include embryonic development, neuronal functions, hematopoiesis, and pathological situations, like wound healing and malignant transformation. The mechanism transmitting extracellular signals ultimately starts with binding of the growth factor to a cell surface receptor, that in many cases carries an intrinsic tyrosine kinase activity (1). These receptors fall into several subgroups sharing structural and functional characteristics. Each subgroup of receptors specifically recognizes a family of structurally homologous growth factors. Perhaps the most striking multiplicity of related growth factors is exemplified by the epidermal growth factor (EGF) 1 family of molecules (2). This six cysteine-containing motif of 45-60 amino acids is shared by all members of the family, and it functions as the receptor binding portion of the molecule. Currently there are four known receptors for EGF-like ligands, constituting the ErbB subgroup of receptor tyrosine kinases (also known as HER, or type I receptor tyrosine kinases (3)). Whereas ErbB-1 binds many ligands, including EGF, transforming growth factor ␣ (TGF␣), and amphiregulin, both ErbB-3 and ErbB-4 bind to a family of isoforms, collectively known as neuregulins (also called Neu differentiation factors, heregulins, glial growth factors, and acetylcholine receptor inducing activity) (4). A related group of molecules, termed NRG2, binds to the same two receptors (5-7), and a third molecule, NRG3, exclusively binds to ErbB-4 (8). Two other ligands, betacellulin (9), and the heparin-binding EGF-like growth factor (10, 11) bind to both ErbB-1 and ErbB-4. Interestingly, the m...

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Detection of ERK activation by a novel monoclonal antibody

Cited by 130 publications

References 25 publications

Thrombospondin-1 differentially induces chemotaxis and DNA synthesis of human venous smooth muscle cells at the receptor-binding level

Thrombospondin-1 differentially induces chemotaxis and DNA synthesis of human venous smooth muscle cells at the receptor-binding level

Neuregulin-4: a novel growth factor that acts through the ErbB-4 receptor tyrosine kinase

Epiregulin Is a Potent Pan-ErbB Ligand That Preferentially Activates Heterodimeric Receptor Complexes

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