'We developed a sensitive EGF receptor detection method for frozen tissue sections using biotinylated EGF as the primary reagent. The method was directly compared with an immunohistochemical technique based on an anti-EGF receptor monoclonal antibody (MAb EGFR1) in normal human and rat tissues and in human tumors. The method was ... More
PI3P signaling regulates receptor sorting but not transport in the endosomal pathway.
AuthorsPetiot A, Faure J, Stenmark H, Gruenberg J
JournalJ Cell Biol
PubMed ID12975344
'While evidence is accumulating that phosphoinositide signaling plays a crucial role in growth factor and hormone receptor down-regulation, this signaling pathway has also been proposed to regulate endosomal membrane transport and multivesicular endosome biogenesis. Here, we have followed the fate of the down-regulated EGF receptor (EGFR) and bulk transport (fluid ... More
Retention of biologic activity of human epidermal growth factor following conjugation to a blood-brain barrier drug delivery vector via an extended poly(ethylene glycol) linker.
AuthorsDeguchi Y, Kurihara A, Pardridge WM
JournalBioconjug Chem
PubMed ID9893961
'Human brain gliomas overexpress the receptor for epidermal growth factor (EGF), and radiolabeled EGF is a potential peptide radiopharmaceutical for imaging human brain tumors, should this peptide be made transportable through the blood-brain barrier (BBB) in vivo. Peptide drug delivery to the brain may be facilitated by conjugating peptide radiopharmaceuticals ... More
In Alzheimer's disease the Golgi apparatus of a population of neurons without neurofibrillary tangles is fragmented and atrophic.
AuthorsStieber A, Mourelatos Z, Gonatas NK
JournalAm J Pathol
PubMed ID8579105
'Recent immunocytochemical and morphometric studies in amyotrophic lateral sclerosis, Alzheimer's disease (AD), and aging indicate that the neuronal Golgi apparatus is a reliable index of activity or degeneration. To further evaluate a possible role of the Golgi apparatus in the pathogenesis of AD, we examined by double labeling the neuronal ... More
Targeted cellular delivery of quantum dots loaded on and in biotinylated liposomes.
AuthorsSigot V, Arndt-Jovin DJ, Jovin TM,
JournalBioconjug Chem
PubMed ID20715851
'We describe the preparation, biophysical characterization, and receptor-mediated cellular internalization of biotinylated lipid particles (BLPs) loaded on the surface and internally with two distinct (colors) of quantum dot (QD) probes. BLPs were formulated with 1.4 and 2.7 mol % PEG-lipids containing either a fusogenic or pH-sensitive lipid to promote bilayer ... More
Optimization of cell surface binding enhances efficiency and specificity of molecular conjugate gene delivery.
AuthorsSchaffer DV, Lauffenburger DA
JournalJ Biol Chem
PubMed ID9774415
'Molecular conjugates, or polyplexes, are promising synthetic vectors for targeted, in vivo gene delivery, if their efficiency can be improved. Gaining mechanistic information on conjugate gene delivery can potentially yield significant improvements in transfer efficiency by revealing barriers to conjugate transfer from the cell surface to the nucleus. We have ... More
Green- and red-fluorescent nanospheres for the detection of cell surface receptors by flow cytometry.
AuthorsBhalgat MK, Haugland RP, Pollack JS, Swan S, Haugland RP
JournalJ Immunol Methods
PubMed ID9831388
'Fluorescent probes serve as sensitive tools for obtaining structural and functional information in cellular systems. In spite of the high sensitivity provided by fluorescent reagents, cell surface receptors expressed in low numbers often escape detection with commonly used fluorescent probes. R-Phycoerythrin (R-PE), a molecule with a very high quantum yield, ... 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
Localization and quantification of epidermal growth factor receptors on single cells by confocal laser scanning microscopy.
AuthorsGood MJ, Hage WJ, Mummery CL, De Laat SW, Boonstra J
JournalJ Histochem Cytochem
PubMed ID1506672
We have established a method for quantifying binding of fluorescence-labeled growth factors to their receptors on single cells in situ with the confocal laser scanning microscope (CLSM). Biotinylated epidermal growth factor (EGF) coupled to phycoerythrin-labeled anti-biotin was used to compare the levels of fluorescence on three different cell types for ... More
Dual mode of action of a human anti-epidermal growth factor receptor monoclonal antibody for cancer therapy.
AuthorsBleeker WK, Lammerts van Bueren JJ, van Ojik HH, Gerritsen AF, Pluyter M, Houtkamp M, Halk E, Goldstein J, Schuurman J, van Dijk MA, van de Winkel JG, Parren PW,
JournalJ Immunol
PubMed ID15383606
Epidermal growth factor receptor (EGF-R) overexpression is common in a large number of solid tumors and represents a negative prognostic indicator. Overexpression of EGF-R is strongly tumor associated, and this tyrosine kinase type receptor is considered an attractive target for Ab therapy. In this study, we describe the evaluation of ... More
Coordination between the actin cytoskeleton and membrane deformation by a novel membrane tubulation domain of PCH proteins is involved in endocytosis.
AuthorsTsujita K, Suetsugu S, Sasaki N, Furutani M, Oikawa T, Takenawa T,
JournalJ Cell Biol
PubMed ID16418535
The conserved FER-CIP4 homology (FCH) domain is found in the pombe Cdc15 homology (PCH) protein family members, including formin-binding protein 17 (FBP17). However, the amino acid sequence homology extends beyond the FCH domain. We have termed this region the extended FC (EFC) domain. We found that FBP17 coordinated membrane deformation ... More
Dynein is required for receptor sorting and the morphogenesis of early endosomes.
AuthorsDriskell OJ, Mironov A, Allan VJ, Woodman PG
JournalNat Cell Biol
PubMed ID17173037
The early endosome is organised into domains to ensure the separation of cargo. Activated mitogenic receptors, such as epidermal growth factor (EGF) receptor, are concentrated into vacuoles enriched for the small GTPase Rab5, which progressively exclude nutrient receptors, such as transferrin receptor, into neighbouring tubules. These vacuoles become enlarged, increase ... More
Large-scale screening assay to measure epidermal growth factor internalization.
Authorsde Wit R, Hendrix CM, Boonstra J, Verkleu AJ, Post JA
JournalJ Biomol Screen
PubMed ID10894756
Recently, we showed that the internalization of the epidermal growth factor (EGF) receptor is inhibited by hydrogen peroxide (H(2)O(2)) in human fibroblasts. In order to test the effect of various stress conditions on receptor internalization and to test a variety of antioxidants in their capacity to prevent or reduce the ... More
Quantum dot ligands provide new insights into erbB/HER receptor-mediated signal transduction.
AuthorsLidke DS, Nagy P, Heintzmann R, Arndt-Jovin DJ, Post JN, Grecco HE, Jares-Erijman EA, Jovin TM
JournalNat Biotechnol
PubMed ID14704683
The erbB/HER family of transmembrane receptor tyrosine kinases (RTKs) mediate cellular responses to epidermal growth factor (EGF) and related ligands. We have imaged the early stages of RTK-dependent signaling in living cells using: (i) stable expression of erbB1/2/3 fused with visible fluorescent proteins (VFPs), (ii) fluorescent quantum dots (QDs) bearing ... More
Automated quantification of quantum-dot-labelled epidermal growth factor receptor internalization via multiscale image segmentation.
AuthorsKriete A, Papazoglou E, Edrissi B, Pais H, Pourrezaei K
JournalJ Microsc
PubMed ID16734710
The ability to monitor epidermal growth factor receptor (EGFr) internalization specifically, and cellular protein concentrations and activation states in general, has been recently improved by the use of appropriately functionalized quantum dots (QDs), as a result of the long-lasting fluorescence, brightness and multicolour of these nanoparticles. However, important quantitative information ... More
A novel fluorogenic substrate for detecting alkaline phosphatase activity in situ.
AuthorsHuang Z, You W, Haugland RP, Paragas VB, Olson NA, Haugland RP
JournalJ Histochem Cytochem
PubMed ID8419466
We describe here the in situ detection of alkaline phosphatase (APase) activity with a new fluorogenic substrate, 2-(5'-chloro-2'-phosphoryloxyphenyl)-6-chloro-4-(3H)-quinazolinone (CPPCQ). CPPCQ is very soluble and colorless. APase converts it into a rapidly precipitating product, whose strong fluorescence marks the sites of APase activity. The detected APase was either a probing enzyme ... More
Electric field-induced redistribution and postfield relaxation of epidermal growth factor receptors on A431 cells.
AuthorsGiugni TD, Braslau DL, Haigler HT
JournalJ Cell Biol
PubMed ID3494733
The lateral mobility of the epidermal growth factor (EGF) receptor in the plane of the plasma membrane of cultured A431 cells was investigated using direct and indirect fluorescent probes to measure the generation and relaxation of electric field-induced receptor asymmetry. A steady electric field of 15 V/cm for 30 min ... More
AP-2/Eps15 interaction is required for receptor-mediated endocytosis.
AuthorsBenmerah A, Lamaze C, Bègue B, Schmid SL, Dautry-Varsat A, Cerf-Bensussan N
JournalJ Cell Biol
PubMed ID9490719
We have previously shown that the protein Eps15 is constitutively associated with the plasma membrane adaptor complex, AP-2, suggesting its possible role in endocytosis. To explore the role of Eps15 and the function of AP-2/Eps15 association in endocytosis, the Eps15 binding domain for AP-2 was precisely delineated. The entire COOH-terminal ... More
Epidermal growth factor radiopharmaceuticals: 111In chelation, conjugation to a blood-brain barrier delivery vector via a biotin-polyethylene linker, pharmacokinetics, and in vivo imaging of experimental brain tumors.
AuthorsKurihara A, Deguchi Y, Pardridge WM
JournalBioconjug Chem
PubMed ID10346884
Epidermal growth factor (EGF) is a potential peptide radiopharmaceutical for detection of brain tumors, because many human gliomas overexpress the EGF receptor (EGFR). The transport of EGF to the brain, however, is restricted by the blood-brain barrier (BBB). The purpose of the present study was to develop a vector-mediated brain ... More
Crosstalk between CLCb/Dyn1-Mediated Adaptive Clathrin-Mediated Endocytosis and Epidermal Growth Factor Receptor Signaling Increases Metastasis.
Authors
JournalDev Cell
PubMed ID28171750
Chloroquine inhibits autophagic flux by decreasing autophagosome-lysosome fusion.
Authors
JournalAutophagy
PubMed ID29940786
The N-recognin UBR4 of the N-end rule pathway is targeted to and required for the biogenesis of the early endosome.
Authors
JournalJ Cell Sci
PubMed ID30111582
Spatiotemporal three-dimensional transport dynamics of endocytic cargos and their physical regulations in cells.
Authors
JournaliScience
PubMed ID35479412
FCHSD2 controls oncogenic ERK1/2 signaling outcome by regulating endocytic trafficking.
Authors
JournalPLoS Biol
PubMed ID32678845
EGFR transactivates RON to drive oncogenic crosstalk.
Authors
JournalElife
PubMed ID34821550
EGFR Expression in HER2-Driven Breast Cancer Cells.
Authors
JournalInt J Mol Sci
PubMed ID33260837
Beclin 1 regulates growth factor receptor signaling in breast cancer.