scholarly journals ImpairedO-LinkedN-Acetylglucosaminylation in the Endoplasmic Reticulum by Mutated Epidermal Growth Factor (EGF) Domain-specificO-LinkedN-Acetylglucosamine Transferase Found in Adams-Oliver Syndrome

2014 ◽  
Vol 290 (4) ◽  
pp. 2137-2149 ◽  
Author(s):  
Mitsutaka Ogawa ◽  
Shogo Sawaguchi ◽  
Takami Kawai ◽  
Daita Nadano ◽  
Tsukasa Matsuda ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Epidermal Growth ◽  
Egf Domain ◽  
Acetylglucosamine Transferase

Orientation of the v-erb-B gene product in the plasma membrane

1986 ◽  
Vol 6 (4) ◽  
pp. 1329-1333
Author(s):  
R C Schatzman ◽  
G I Evan ◽  
M L Privalsky ◽  
J M Bishop
Keyword(s):  
Endoplasmic Reticulum ◽  
Epidermal Growth Factor Receptor ◽  
Plasma Membrane ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Growth Factor Receptor ◽  
Kinase Domain ◽  
Amino Terminus ◽  
Protein Kinase Domain ◽  
Epidermal Growth

The retroviral oncogene v-erb-B encodes a truncated version of the receptor for epidermal growth factor. To define the disposition of the v-erb-B protein within cells and across the plasma membrane, we raised antibodies against defined epitopes in the protein and used these in immunofluorescence to analyze cells transformed by v-erb-B. A small fraction of the v-erb-B protein was found on the plasma membrane in a clustered configuration. The bulk of the protein was located in the endoplasmic reticulum and Golgi apparatus. Epitopes near the amino terminus of the v-erb-B protein were displayed on the surface of the cell, whereas epitopes in the protein kinase domain were located exclusively within cells. We conclude that the v-erb-B protein spans the plasma membrane in a manner similar or identical to that of the epidermal growth factor receptor, even though the viral transforming protein does not possess the signal peptide that is thought to direct insertion of the receptor into the membrane.

Mechanism regulating nuclear calcium signalingThis paper is one of a selection of papers published in this Special Issue, entitled The Nucleus: A Cell Within A Cell.

2006 ◽  
Vol 84 (3-4) ◽  
pp. 403-422 ◽  
Author(s):  
Anant N. Malviya ◽  
Christian Klein
Keyword(s):  
Endoplasmic Reticulum ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Calcium Signaling ◽  
Nuclear Membrane ◽  
Protein Fragment ◽  
Outer Nuclear Membrane ◽  
Nuclear Calcium ◽  
A Cell ◽  
Epidermal Growth

Although the outer nuclear membrane is continuous with the endoplasmic reticulum, it is possible to isolate nuclei both intact and free from endoplasmic reticulum contaminants. The outer and the inner nuclear membranes can be purified free from cross-contamination. Evidence in support of autonomous regulation of nuclear calcium signaling relies upon the investigations with isolated nuclei. Mechanisms for generating calcium signaling in the nucleus have been identified. Two calcium transporting systems, an ATP-dependant nuclear Ca2+-ATPase and an IP4-mediated inositol 1,3,4,5-tetrakisphosphate receptor, are located on the outer nuclear membrane. Thus, ATP and IP4, depending on external free calcium concentrations, are responsible for filling the nuclear envelope calcium pool. The inositol 1,4,5-trisphosphate receptor is located on the inner nuclear membrane with its ligand binding domain facing toward the nucleoplasm. Likewise, the ryanodine receptor is located on the inner nuclear membrane and its ligand cADP-ribose is generated within the nucleus. A 120 kDa protein fragment of nuclear PLC-γ1 is stimulated in vivo by epidermal growth factor nuclear signaling coincident with the time course of nuclear membrane epidermal growth factor receptor activation. Stimulated 120 kDa protein fragment interacts with PIKE, a nuclear GTPase, and together they form a complex with PI[3]kinase serving as a module for nuclear PI[3]K stimulation. Thus, the nucleus has its own IP3 generating system.

scholarly journals The effect of epidermal growth factor (EGF) on the differentiation of the rough endoplasmic reticulum in fetal mouse small intestine in organ culture.

1981 ◽  
Vol 29 (6) ◽  
pp. 765-770 ◽  
Author(s):  
J F Beaulieu ◽  
R Calvert
Keyword(s):  
Endoplasmic Reticulum ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Organ Culture ◽  
Rough Endoplasmic Reticulum ◽  
Fetal Mouse ◽  
Absorptive Cells ◽  
Mouse Small Intestine ◽  
Epidermal Growth ◽  
Mouse Fetuses

The differentiation of the rough endoplasmic reticulum (RER) of mouse duodenal absorptive cells located at the tip of the villi at 17 days of gestation was compared to that of absorptive cells in duodenal explants of 15-day-old mouse fetuses cultured for 72 hr 1) with Trowell T8 medium (without insulin) alone or supplemented 2) with epidermal growth factor (EGF; 100 ng/ml) or 3) with 25% bovine amniotic fluid (BAF). Glucose-6-phosphatase activity (G6Pase) was localized cytochemically to ensure a better identification of the RER. The intersections of a double lattice falling over and outside the RER were counted and the percentage of intersections over the RER was estimated. With this method, the extent of the RER is not statistically different when the absorptive cells in utero are compared to those of explants cultured with EGF. However, the extent of the RER in the absorptive cells cultured with Trowell T8 medium alone or supplemented with BAF is 50% lower than in the former two groups. It is concluded that EGF promotes the maturation of duodenal absorptive cells in organ culture.

Processing and targeting of proteins in the eucaryote

1988 ◽  
Vol 66 (12) ◽  
pp. 1253-1257 ◽  
Author(s):  
J. J. M. Bergeron
Keyword(s):  
Endoplasmic Reticulum ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Golgi Apparatus ◽  
Tyrosine Kinases ◽  
Receptor Tyrosine Kinases ◽  
Endoplasmic Reticulum Membrane ◽  
Extracellular Milieu ◽  
Epidermal Growth ◽  
Selective Processing

Cell-free systems have helped to elucidate the molecular constituents regulating the selection and translocation of proteins into and across the endoplasmic reticulum membrane, chloroplasts, mitochondria, and peroxisomes, the transport to and through the Golgi apparatus, and the sorting of proteins to the lysosomes, the plasmalemma, and the extracellular milieu. The use of cell-free systems has also been instrumental in defining the endosomal apparatus and its functional significance in the sorting of incoming ligands and receptors, the selective processing of internalized ligands such as insulin, and transmembrane signalling, especially of the epidermal growth factor and insulin receptor tyrosine kinases. Predicted use of cell-free systems to study interorganelle relationships may help to identify the majority of molecular constituents regulating membrane traffic in the eucaryote.

311 ULTRASTRUCTURAL ANALYSIS OF OVINE PREANTRAL FOLLICLES CULTURED IN THE PRESENCE OF INDOLE ACETIC ACID, EGF, AND FSH

2006 ◽  
Vol 18 (2) ◽  
pp. 263
Author(s):  
E. Andrade ◽  
F. Landim-Alvarenga ◽  
J. Silva ◽  
M. Max ◽  
A. Alfieri ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Acetic Acid ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Indole Acetic Acid ◽  
Primordial Follicles ◽  
Preantral Follicles ◽  
Epidermal Growth ◽  
Cortical Slices

Previous studies from our team demonstrated that ovine primordial follicles are successfully activated in vitro after culturing in medium supplemented with 40 ng/mL indole acetic acid (IAA); besides that, the addition of IAA and epidermal growth factor (EGF) or EGF and follicle stimulating hormone (FSH) to the culture media were the most effective treatments to sustain the health and viability of activated ovine primordial follicles during in vitro culture. In this work, follicular quality was assessed only by histological studies; there is a great need to evaluate ultrastructural changes occurring in primordial follicles during activation in vitro. The aim of this study was to investigate the ultrastructural characteristics of preantral follicles after culturing of cortical slices in media containing IAA, EGF, and FSH, alone and in combination. Ovaries (n = 8) from adult merino ewes were collected at local slaughterhouses. Small pieces of ovarian fragments were removed for transmission electron microscopy (TEM). These pieces of ovarian cortex were cultured in culture dishes containing 1 mL aliquots of culture medium at 39�C with 5% CO2 in air. The media used were: (T1) Minimum essential medium (MEM) supplemented with ITS (insulin 6.25 �g/mL, transferrin 6.25 �g/mL and selenium 6.25 ng/mL), 0.23 mM pyruvate, 2 mM glutamine, 2 mM hypoxantine, 1.25 mg/mL bovine serum albumin (BSA), and antibiotics (100 ��g/mL penicillin and 100 ��g/mL streptomycim) (MEM+, control medium); (T2) MEM+IAA (40 ng/mL); (T3) MEM+IAA + epidermal growth factor (EGF) (100 ng/mL; Sigma, St. Louis, MO, USA); or (T4) MEM+EGF+FSH (100 ng/mL). After 6 days of culture of cortex tissue fragments in media, ultrastructural analysis was performed on preantral follicles (n = 3 in each treatment) included in a small cortical fragments. Preantral follicles were classified according to the stage of development as primordial follicles or as developing follicles. Preantral follicles cultured in supplemented media for 6 days were ultrastructurally normal. Their oocytes had an intact nucleus and cytoplasm that contained heterogeneous-sized lipid droplets, and numerous round or elongated mitochondria with intact parallel cristae were observed. Occasionally these organelles were associated with smooth endoplasmic reticulum (SER). Rough endoplasmic reticulum (RER) was rarely found. The cytoplasm of granulosa cells contained a large number of mitochondria and abundant RER. In contrast, follicles cultured in MEM+ (control) had a large number of vacuoles in the oocyte cytoplasm and excessive clustering of the chromatin material in the nucleus, suggesting an initial process of oocyte degeneration. In conclusion, the presence of IAA, EGF, FSH and their combinations helped to maintain ultrastructural integrity of ovine preantral follicles in cortical slices cultured in vitro.

Production of a human epidermal growth factor fusion protein and its degradation in rat gastrointestinal flushings

1996 ◽  
Vol 16 (1) ◽  
pp. 89-97 ◽  
Author(s):  
C J Xian ◽  
Z Upton ◽  
C Goddard ◽  
C A Shoubridge ◽  
K A McNeil ◽  
...  
Keyword(s):  
Biological Activity ◽  
Growth Factor ◽  
Small Bowel ◽  
Epithelial Cell ◽  
Epidermal Growth Factor ◽  
Fusion Protein ◽  
Egf Receptor ◽  
Human Epidermal Growth Factor ◽  
Epidermal Growth ◽  
Egf Domain

ABSTRACT This study describes the biosynthesis of a human epidermal growth factor fusion protein, Long EGF, that has a 53 amino acid extension peptide derived from the 46 N-terminal amino acids of porcine GH. The approach allowed the production of Long EGF at high efficiency due to the expression of the fusion protein in high yield as inclusion bodies in Escherichia coli. Long EGF had a slightly lower potency compared with native EGF in a range of assays, including binding to anti-EGF antibodies or the EGF receptor, stimulation of Balb/3T3 fibroblast and rat intestinal epithelial cell growth, as well as counteracting the inhibition of mink lung epithelial cell proliferation by transforming growth factor-β1. Degradation of Long EGF and native EGF was compared in gastrointestinal flushings as an indication of whether the EGF domain of the fusion protein would be protected from proteolytic cleavage and be useful as a trophic agent in the gut. Incubation with flushings from the stomach or jejunum of rats caused rapid cleavage of the extension peptide, releasing native EGF. A C-terminal truncation of Arg53 in the stomach and a removal of the C-terminal pentapeptide (49Trp-Trp-Glu-Leu-Arg53) in the small bowel was demonstrated by N-terminal sequencing and mass spectrometry. The degradation patterns were reflected by changes in migration of products on SDS-PAGE and in subsequent binding activities to the EGF receptor and anti-EGF antibodies. The data show that a human EGF fusion protein can be produced efficiently in a bacterial expression system and that it retains biological activity in vitro. Although the extension peptide was rapidly cleaved from Long EGF in both stomach and small bowel producing similar biological activity to native EGF, it could not prevent subsequent degradation of the EGF domain. Other strategies are being investigated to develop an effective oral form of EGF that resists digestion by proteases in the gastrointestinal tract.

scholarly journals O-GlcNAcylation of the human epidermal growth factor receptor

2015 ◽  
Vol 13 (30) ◽  
pp. 8196-8204 ◽  
Author(s):  
Silviya R. Stateva ◽  
Antonio Villalobo
Keyword(s):  
Epidermal Growth Factor Receptor ◽  
Plasma Membrane ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Protein Kinases ◽  
Protein Phosphatases ◽  
Growth Factor Receptor ◽  
Human Epidermal Growth Factor ◽  
Epidermal Growth ◽  
Acetylglucosamine Transferase

The cartoon represents the EGFR at the plasma membrane where serine/threonine residues could be subjected to phosphorylation/dephosphorylation events by protein kinases (PK) and phospho-protein phosphatases (PPP) and to O-GlcNAcylation/deGlcNAcylation events by O-linked β-N-acetylglucosamine transferase (OGT) and O-linked β-N-acetylglucosaminidase (OGA).

scholarly journals Critical cysteine residues for regulation of integrin alphaIIbbeta3 are clustered in the epidermal growth factor domains of the beta3 subunit

2004 ◽  
Vol 378 (3) ◽  
pp. 1079-1082 ◽  
Author(s):  
Tetsuji KAMATA ◽  
Hironobu AMBO ◽  
Wilma PUZON-McLAUGHLIN ◽  
Kenneth Khiem TIEU ◽  
Makoto HANDA ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Cysteine Residue ◽  
Cysteine Residues ◽  
Disulphide Bonds ◽  
Binding Function ◽  
Beta3 Subunit ◽  
Epidermal Growth ◽  
Egf Domain

Chemical or enzymic reduction/oxidation of integrin cysteine residues (e.g. by reducing agents and protein disulphide isomerase) may be a mechanism for regulating integrin function. It has also been proposed that unique cysteine residues in the integrin β3 subunit are involved in the regulation of αIIbβ3. In the present study, we studied systematically the role of disulphide bonds in β3 on the ligand-binding function of αIIbβ3 by mutating individual cysteine residues of β3 to serine. We found that the disulphide bonds that are critical for αIIbβ3 regulation are clustered within the EGF (epidermal growth factor) domains. Interestingly, disrupting only a single disulphide bond in the EGF domains was enough to activate αIIbβ3 fully. In contrast, only two (of 13) disulphide bonds tested outside the EGF domains activated αIIbβ3. These results suggest that the disulphide bonds in the EGF domains should be intact to keep αIIbβ3 in an inactive state, and that there is no unique cysteine residue in the EGF domain critical for regulating the receptor. The cysteine residues in the EGF domains are potential targets for chemical or enzymic reduction.

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