Binding, sequestration, and processing of epidermal growth factor and nerve growth facor by PC12 cells

Chandler, Charles F.; Herschman, Harvey R.

doi:10.1002/jcp.1041140311

Cited by 26 publications

(13 citation statements)

References 35 publications

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“…EGF Receptors-In addition to mAChRs, PC12 cells also express membrane receptors for NGF and EGF (20,75). It has been reported that EGF receptors can be transactivated by mAChR agonists via extracellular and intracellular signal pathways (76 -79).…”

Section: Activation Of Rap1 By Carbachol Does Not Occur Via Ngf Ormentioning

confidence: 99%

A CalDAG-GEFI/Rap1/B-Raf Cassette Couples M1Muscarinic Acetylcholine Receptors to the Activation of ERK1/2

Guo

Kumahara

Saffen

2001

Journal of Biological Chemistry

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In this study we examine signaling pathways linking the M 1 subtype of muscarinic acetylcholine receptor (M 1 mAChR) to activation of extracellular signal-regulated kinases (ERK) 1 and 2 in neuronal PC12D cells. We first show that activation of ERK1/2 by the M 1 mAChR agonist carbachol takes place primarily via a Ras-independent pathway that depends largely upon Rap1, another small GTP-binding protein in the Ras family. Rap1 in turn activates B-Raf, an upstream activator of ERK1/2. Consistent with these results, carbachol was found to activate Rap1 more potently than Ras. Similar to other small GTP-binding proteins, activation of Rap1 requires a guanine nucleotide exchange factor (GEF) to promote its conversion from the GDP-to GTP-bound form. Using specific antibodies, we show that a recently identified Rap1 GEF, calcium-and diacylglycerol-regulated guanine nucleotide exchange factor I (CalDAG-GEFI), is expressed in PC12D cells and that carbachol stimulates the formation of a complex containing CalDAG-GEFI, Rap1, and activated B-Raf. Finally, we show that expression of CalDAG-GEFI antisense RNA largely blocks carbachol-stimulated activation of hemagglutinin (HA)1-tagged B-Raf and formation of the CalDAG-GEFI/Rap1/ HA1-tagged B-Raf complex. Together, these data define a novel signaling pathway for M 1 mAChR, where increases in Ca 2؉ and diacylglycerol stimulate the sequential activation of CalDAG-GEFI, Rap1, and B-Raf, resulting in the activation of MEK and ERK1/2.

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Section: Activation Of Rap1 By Carbachol Does Not Occur Via Ngf Ormentioning

confidence: 99%

A CalDAG-GEFI/Rap1/B-Raf Cassette Couples M1Muscarinic Acetylcholine Receptors to the Activation of ERK1/2

Guo

Kumahara

Saffen

2001

Journal of Biological Chemistry

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“…Furthermore, these results suggest a large potential for hidden NGF might exist in trk A-bearing structures such as NGF-responsike nerve processes. This detergent in the homogenization buffer and includes a proposal is further strengthened by the wealth of evi-dence that indicates NGF forms a stable complex with this receptor after acquisition (Calissano and Shelanski, 1980;Olender and Stach, 1980;Olender et al 1981;Chandler and Herschman, 1983;Puma et al, 1983;Bernd and Greene, 1984;Kasaian and Neet, 1988).…”

Section: Evidence For Hidden Ngf In Tissue Extractsmentioning

confidence: 99%

Detection of increased tissue concentrations of nerve growth factor with an improved extraction procedure

et al. 1996

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Nerve growth factor (NGF) is a protein essential for the survival and normal function of sympathetic neurons. Two‐site immunoassays have been developed over the past decade in several laboratories and used to estimate its endogenous concentrations in a variety of effector tissues. However, levels appear restricted to a narrow range, display only a poor correlation with innervation density, and show obvious inter‐ and intralaboratory variations, the origins of which are unclear. This led us to examine alternative extraction procedures for NGF before quantification. In particular, we have found treatment of tissue extracts with high and low pH in the presence of detergent results in the detection of higher NGF concentrations in immunoassays using either polyclonal or commercially available monoclonal antibodies. These increases were tissue‐specific (sciatic nerve, mesenteric arteries, and thoracic aorta > heart and brain > sympathetic ganglia > abdominal aorta) and as much as 10 times greater than the amounts detected by traditional procedures. The method should also prove useful for the assay of other members of the neurotrophin family when appropriate antibodies become available. © 1996 Wiley‐Liss, Inc.

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“…These observations are of interest in that EGF and NGF share a set of positive pleiotypic actions on these cells and are processed by the cell in a generally similar manner (29,30).…”

mentioning

confidence: 97%

Nerve growth factor- and epidermal growth factor-stimulated phosphorylation of a PC12 cytoskeletally associated protein in situ.

Landreth

Rieser

1985

The Journal of Cell Biology

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Nerve growth factor (NGF) and epidermal growth factor (EGF) produce stable alterations in PC12 cells that persist in the detergent-insoluble cytoskeleton, resulting in the phosphorylation of a 250,000-mol-wt cytoskeletally associated protein in situ. Treatment of PC12 cells with NGF or EGF, followed by detergent lysis of the cells and incubation of the resulting cytoskeletons with 3,-32p-ATP, permitted detection of hormonally stimulated, energydependent events, which result in the enhanced phosphorylation of a cytoskeletally associated protein as an immediate consequence of receptor occupancy. These events were elicited only upon treatment of intact cells at physiological temperatures. The NGF-and EGFstimulated events occurred rapidly; however, they were a transient effect of hormone action. NGF and EGF were found to act through independent mechanisms to stimulate the in situ phosphorylation of the 250,000-mol-wt protein, as the effects of NGF, but not EGF, were blocked by methyltransferase inhibitors. The 250,000-mol-wt protein was phosphorylated on serine and threonine residues in response to both NGF and EGF although in somewhat different proportions. The data suggest that the hormone-stimulated labeling of the 250,000-mol-wt protein may be the result of either the direct activation of a protein kinase, the redistribution of the kinase relative to its substrates as a consequence of hormone action, or the coincident occurrence of these events.Although it is well known that the binding of peptide hormones to cell surface receptors results in various metabolic changes in the target cells, the mechanisms subserving these events are not well understood. Several hypotheses have been advanced to explain the myriad effects of peptide hormones, including the involvement of second messengers generated on hormone binding (1), internalization of the hormone and subsequent transport to the nucleus (2), and the activation of hormonally sensitive protein kinases (3). There is reasonably good evidence that at least some of the biological effects, including the stimulation of DNA synthesis, are the result of receptor aggregation within the plasma membrane (4-6).The role of protein kinases in hormone action is of particular interest, given the recent discovery that the membrane receptors for several peptide hormones possess an intrinsic protein kinase activity (7-12). The binding of hormone to its receptor immediately activates the protein kinase, resulting in autophosphorylation of the receptor itself as well as the phosphorylation of other cellular proteins (3,7,(12)(13)(14). It is

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Binding, sequestration, and processing of epidermal growth factor and nerve growth facor by PC12 cells

Cited by 26 publications

References 35 publications

A CalDAG-GEFI/Rap1/B-Raf Cassette Couples M1Muscarinic Acetylcholine Receptors to the Activation of ERK1/2

A CalDAG-GEFI/Rap1/B-Raf Cassette Couples M1Muscarinic Acetylcholine Receptors to the Activation of ERK1/2

Detection of increased tissue concentrations of nerve growth factor with an improved extraction procedure

Nerve growth factor- and epidermal growth factor-stimulated phosphorylation of a PC12 cytoskeletally associated protein in situ.

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