'Treatment of Swiss mouse 3T3 fibroblasts with certain cyclic nucleotide phosphodiesterase inhibitors (theophylline, SQ 20,006, and MY-5445) prevents the activation of the M(r) 70,000 S6 kinase (p70S6k) induced by a variety of external stimuli. Concentrations giving half-maximal inhibition were 800, 50, and 25 microM, respectively. Western blot analysis and immunocomplex ... More
Biotin-ligand complexes with streptavidin quantum dots for in vivo cell labeling of membrane receptors.
AuthorsLidke DS, Nagy P, Jovin TM, Arndt-Jovin DJ,
JournalMethods Mol Biol
PubMed ID17237530
'The unique fluorescence properties of quantum dots (QDs), particularly their large extinction coefficients and photostability, make them ideal probes for tracking proteins in live cells using real-time visualization. We have shown that QDs conjugated to epidermal growth factor act as functional ligands for their receptor, erbB1. Here, we describe protocols ... More
Reaching out for signals: filopodia sense EGF and respond by directed retrograde transport of activated receptors.
AuthorsLidke DS, Lidke KA, Rieger B, Jovin TM, Arndt-Jovin DJ
JournalJ Cell Biol
PubMed ID16103229
'ErbB1 receptors situated on cellular filopodia undergo systematic retrograde transport after binding of the epidermal growth factor (EGF) and activation of the receptor tyrosine kinase. Specific inhibitors of the erbB1 receptor tyrosine kinase as well as cytochalasin D, a disruptor of the actin cytoskeleton, abolish transport but not free diffusion ... More
Nonradioactive ligand binding assay for epidermal growth factor receptor.
AuthorsKing IC, Catino JJ
JournalAnal Biochem
PubMed ID2221374
'A rapid and sensitive nonradioactive ligand binding assay for identifying the agonists and antagonists of epidermal growth factor receptor is described. The assay is easy to perform, has a within-assay error of less than 15%, and does not use radioactive substances. Nonspecific binding is less than 10% and the assay ... More
Direct linkage of EGF to its receptor: characterization and biological relevance.
AuthorsLinsley PS, Fox CF
JournalJ Supramol Struct
PubMed ID6264232
'A small portion of the 125I-EGF that binds specifically to intact cells or isolated membrane from a variety of sources becomes directly and irreversibly linked to EGF receptors. This provides a simple technique for affinity labeling the EGF receptor. Membranes isolated from the human epidermoid carcinoma cell line A431, which ... More
Survival factor-insensitive generation of reactive oxygen species induced by serum deprivation in neuronal cells.
AuthorsSatoh T, Sakai N, Enokido Y, Uchiyama Y, Hatanaka H
JournalBrain Res
PubMed ID8891242
'To investigate the involvement of reactive oxygen species (ROS) in neuronal apoptosis, we performed confocal and flow cytometric analysis with a ROS-specific fluorogen, 6-carboxy-2'', 7''-dichorodihydrofluorescein diacetate, di(acetoxymethyl ester) (C-DCDHF-DA). Serum deprivation significantly increased the level of ROS in PC12 cells and rat cortical neurons. N,N''-diphenyl-p-phenylenediamine (DPPD), an antioxidant, reduced ROS ... More
Receptors for epidermal growth factor and other polypeptide mitogens.
AuthorsCarpenter G
JournalAnnu Rev Biochem
PubMed ID3039909
Nobel lecture. Epidermal growth factor.
AuthorsCohen S
JournalBiosci Rep
PubMed ID3555641
A biological assay for epidermal growth factor/urogastrone and related polypeptides.
AuthorsCarpenter G, Zendegui J
JournalAnal Biochem
PubMed ID2422978
A bioassay for epidermal growth factor (EFG) which is rapid, specific, and extremely sensitive is described. As the bioassay detects as little as 25 pg of EGF, this assay is more sensitive than commonly used radioreceptor assays and nearly as sensitive as radioimmunoassays. This bioassay involves only the measurement of ... More
Rotational mobility of high-affinity epidermal growth factor receptors on the surface of living A431 cells.
AuthorsZidovetzki R, Johnson DA, Arndt-Jovin DJ, Jovin TM
JournalBiochemistry
PubMed ID1647816
The rotational diffusion of epidermal growth factor (EGF) bound to its specific receptor on the surface of human carcinoma A431 cells was studied by means of time-resolved phosphorescence anisotropy measurements. The rotational mobility was measured on the total population of EGF receptors by using a saturating concentration of EGF conjugated ... More
Epidermal growth factor receptors.
AuthorsAdamson ED, Rees AR
JournalMol Cell Biochem
PubMed ID6261114
EGF-Rs are cell membrane glycoproteins of wide distribution. They have not yet been fully characterized or purified but are probably molecules of 170-190,000 mol. wt. in most cells. The growth factor EGF binds and will saturate cell surface receptors with a KA of about 5 X 10(9) M-1 although a ... More
Recycling of epidermal growth factor-receptor complexes in A431 cells: identification of dual pathways.
AuthorsSorkin A, Krolenko S, Kudrjavtceva N, Lazebnik J, Teslenko L, Soderquist AM, Nikolsky N
JournalJ Cell Biol
PubMed ID1986007
The intracellular sorting of EGF-receptor complexes (EGF-RC) has been studied in human epidermoid carcinoma A431 cells. Recycling of EGF was found to occur rapidly after internalization at 37 degrees C. The initial rate of EGF recycling was reduced at 18 degrees C. A significant pool of internalized EGF was incapable ... More
Real-time measurements of kinetics of EGF binding to soluble EGF receptor monomers and dimers support the dimerization model for receptor activation.
AuthorsZhou M, Felder S, Rubinstein M, Hurwitz DR, Ullrich A, Lax I, Schlessinger J
JournalBiochemistry
PubMed ID8347619
We have tested one aspect of the allosteric dimerization model for the activation of EGF receptor (EGFR) by EGF: whether EGF binding favors dimerization of the receptor. For this to be true, EGF molecules must bind with higher affinity to dimeric receptors than to monomeric receptors. We have tested this ... More
Temperature-dependent lateral and transverse distribution of the epidermal growth factor receptor in A431 plasma membranes.
AuthorsAzevedeo JR, Johnson DA
JournalJ Membr Biol
PubMed ID2077131
To elucidate further the structure and molecular dynamics of the epidermal growth factor receptor, temperature-dependent aggregation and extracellular protrusion of the epidermal growth factor receptor in isolated plasma membranes from A431 cells were examined by fluorescence energy-transfer techniques. Epidermal growth factor was labeled at the amino terminus with either fluorescein ... More
Critical functional requirement for the guanidinium group of the arginine 41 side chain of human epidermal growth factor as revealed by mutagenic inactivation and chemical reactivation.
AuthorsEngler DA, Campion SR, Hauser MR, Cook JS, Niyogi SK
JournalJ Biol Chem
PubMed ID1733935
In a preliminary study we demonstrated that the formation of the epidermal growth factor (EGF) receptor-ligand complex requires the participation of the highly conserved arginine 41 side chain of the growth factor peptide (Engler, D.A., Montelione, G.T., and Niyogi, S.K. (1990) FEBS Lett. 271, 47-50). In an attempt to gain ... More
Glia modulate the response of murine cortical neurons to excitotoxicity: glia exacerbate AMPA neurotoxicity.
AuthorsDugan LL, Bruno VM, Amagasu SM, Giffard RG
JournalJ Neurosci
PubMed ID7540679
We have developed "pure" neuronal cultures (< 1% astrocytes) from mouse neocortex to study the effect of glial cells on the response of neurons to injury. Cortical neurons were found to require glial-conditioned medium to survive. Immature neurons, 2-4 d in vitro, deprived of glial-conditioned medium, underwent apoptosis over 48 ... More