Nerve growth factor receptors are preaggregated and immobile on responsive cells

Venkatakrishnan, Gita; McKinnon, Christine A.; Pilapil, Carmencita; Wolf, David E.; Ross, Alonzo H.

doi:10.1021/bi00225a002

Cited by 19 publications

(13 citation statements)

References 52 publications

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“…The clustered arrangement of human C5a receptors expressed on the surface of transfected mouse L-cells ( Figures 2E-2G) suggests that clustering is a native property of these receptor molecules. Finally, receptors for nerve growth factor on responsive cells are found preclustered on responsive cells, providing a precedent for receptor preclustering and immobility on a cell surface (Venkatakrishnan et al 1991).…”

Section: Discussionmentioning

confidence: 96%

Receptors for the Chemoattractants C5a and IL-8 Are Clustered on the Surface of Human Neutrophils

Gray

Hasslen

Ember

et al. 1997

J Histochem Cytochem.

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SUMMARYWe have used high-resolution field emission scanning electron microscopy with backscatter electron imaging to detect immunogold-labeled C5a and interleukin-8 (IL-8) receptors on human blood neutrophils. The receptors were labeled with receptor-specific antibodies in combination with secondary antibody conjugated to immunogold. When neutrophils were isolated in a "nonactivated" state, both of these receptor populations were expressed primarily in clusters on nonprojecting domains of the cell membrane. When these cells were double labeled for C5a and IL-8 receptors, intermixing of these receptor species in a common cluster was not found. When neutrophils were isolated in an "activated" state, by mixing the blood with N -formylmethionyl-leucyl-phenylalanine, the cells were seen to be elongated and ruffled at their anterior pole, but the C5a receptors did not disperse or redistribute on the surface of the peptide-activated cells. Analysis of the distribution of human C5a receptors expressed by transfected mouse L-cell fibroblasts showed the C5a receptors to be clustered, but expressed on nonprojecting and projecting domains of the cell surface. These observations provide new information on the topographical expression of leukocyte receptors involved in directing cell migration.

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Section: Discussionmentioning

confidence: 96%

Receptors for the Chemoattractants C5a and IL-8 Are Clustered on the Surface of Human Neutrophils

Gray

Hasslen

Ember

et al. 1997

J Histochem Cytochem.

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“…These findings led to the hypothesis that p75 and TrkA form a heteromolecular complex, which is the high‐affinity NGF receptor (Hempstead et al, 1991). Biophysical approaches using intact cells (Venkatakrishnan et al, 1991; Wolf et al, 1995; Ross et al, 1996) and co‐immunoprecipitation studies with detergent solubilized receptors (Huber and Chao, 1995; Gargano et al, 1997; Ross et al, 1998) provide good evidence for the p75–TrkA complex.…”

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confidence: 99%

Novel functional interactions between Trk kinase and p75 neurotrophin receptor in neuroblastoma cells

Lachyankar

Condon

Daou

et al. 2002

J of Neuroscience Research

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To understand the functional interactions between the TrkA and p75 nerve growth factor (NGF) receptors, we stably transfected LAN5 neuroblastoma cells with an expression vector for ET-R, a chimeric receptor with the extracellular domain of the epidermal growth factor receptor (EGFR), and the TrkA transmembrane and intracellular domains. EGF activated the ET-R kinase and induced partial differentiation. NGF, which can bind to endogenous p75, did not induce differentiation but enhanced the EGF-induced response, leading to differentiation of almost all cells. A mutated NGF, 3T-NGF, that binds to TrkA but not to p75 did not synergize with EGF. Enhancement of EGF-induced differentiation required at least nanomolar concentrations of NGF, consistent with the low-affinity p75 binding site. EGF may induce a limited number of neuronal cells because it also enhanced apoptosis. Both NGF and a caspase inhibitor reduced apoptosis and, thereby, enhanced differentiation. NGF seems to enhance survival through the phosphatidylinositol-3 kinase (PI3K) pathway. Consistent with this hypothesis, Akt, a downstream effector of the PI3K pathway, was hyperphosphorylated in the presence of EGF+NGF. These results demonstrate that TrkA kinase initiates differentiation, and p75 enhances differentiation by rescuing differentiating cells from apoptosis via the PI3K pathway. Even though both EGF and NGF are required for differentiation of LAN5/ET-R cells, only NGF is required for survival of the differentiated cells. In the absence of NGF, the cells die by an apoptotic mechanism, involving caspase-3. An anti-p75 antibody blocked the survival effect of NGF. Brain-derived neurotrophic factor also enhanced cell survival, indicating that in differentiated cells, NGF acts through the p75 receptor to prevent apoptosis.

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“…Using fluorescence recovery after photobleaching (FRAP), we have measured the lateral diffusion of gp75 in intact cells (46,47,54). Gp75 is highly mobile on cells which lack TrkA and are nonresponsive to NGF.…”

mentioning

confidence: 99%

The neurotrophin receptor, gp75, forms a complex with the receptor tyrosine kinase TrkA.

Ross

Daou

McKinnon

et al. 1996

The Journal of Cell Biology

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Abstract. The high-affinity NGF receptor is thought to be a complex of two receptors, gp75 and the tyrosine kinase TrkA, but direct biochemical evidence for such an association has been lacking. In this report, we demonstrate the existence of such a gp75-TrkA complex by a copatching technique. Gp75 on the surface of intact cells is patched with an anti-gp75 antibody and fluorescent secondary antibody, the cells are then fixed to prevent further antibody-induced redistributions, and the distribution of TrkA is probed with an anti-TrkA antibody and fluorescent secondary antibody. We utilize a baculovirus-insect cell expression system which allows high level expression of wild-type and mutated NGF receptors. TrkA and gp75 copatch in both the absence and presence of NGF. This association is specific, since gp75 does not copatch with other tyrosine kinase receptors, including TrkB, platelet-derived growth factor receptor-~, and Torso (Tor). To determine which domains of TrkA are required for copatching, we used a series of TrkA-Tor chimeric receptors and show that the extracellular domain of TrkA is sufficient for copatching with gp75. A chimeric receptor with TrkA transmembrane and intracellular domains shows partial copatching with gp75. Deletion of the intracellular domain of gp75 decreases but does not eliminate copatching. A point mutation which inactivates the TrkA kinase has no effect on copatching, indicating that this enzymatic activity is not required for association with gp75. Hence, although interactions between the gp75 and TrkA extracellular domains are sufficient for complex formation, interactions involving other receptor domains also play a role.

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Nerve growth factor receptors are preaggregated and immobile on responsive cells

Cited by 19 publications

References 52 publications

Receptors for the Chemoattractants C5a and IL-8 Are Clustered on the Surface of Human Neutrophils

Receptors for the Chemoattractants C5a and IL-8 Are Clustered on the Surface of Human Neutrophils

Novel functional interactions between Trk kinase and p75 neurotrophin receptor in neuroblastoma cells

The neurotrophin receptor, gp75, forms a complex with the receptor tyrosine kinase TrkA.

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