Cloning and characterization of a gene encoding pig epidermal growth factor

1991 ◽  
Vol 6 (1) ◽  
pp. 63-70 ◽  
Author(s):  
J. C. Pascall ◽  
D. S. C. Jones ◽  
S. M. Doel ◽  
J. M. Clements ◽  
M. Hunter ◽  
...  
Keyword(s):  
Amino Acid ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Amino Acid Sequence ◽  
Egf Receptor ◽  
Predicted Amino Acid Sequence ◽  
Recombinant Polypeptide ◽  
Gene Encoding ◽  
Epidermal Growth ◽  
Direct Amplification

ABSTRACT A portion of the pig epidermal growth factor (EGF) gene has been isolated and characterized. The nucleotide sequencies of exons 20 and 21, which encode the EGF region of the precursor protein, show 85% similarity with the human EGF gene sequence. In addition, conservation of the intron—exon boundaries between the two species was generally observed. Although the pig exon 21 appeared to lack a single nucleotide at its 5′ end relative to the human gene, sequences obtained by direct amplification of the genomic DNA around the 5′ end of this exon using the polymerase chain reaction, and from a pig EGF cDNA recombinant isolated from a kidney library, indicated that the deletion was probably a cloning artifact. Comparison of the predicted amino acid sequence of pig EGF with that of EGF from other species, as well as with several other polypeptides which bind to the EGF receptor, indicated conservation of Gly18, Tyr37, Gly39 and Arg41 in addition to all six cysteine residues and Leu47, which are known to be critical for biological activity. A synthetic gene encoding the predicted amino acid sequence of pig EGF was expressed in yeast. The recombinant polypeptide was shown to compete with 125I-labelled mouse EGF for binding to cells and to stimulate DNA synthesis in quiescent monolayers of Swiss 3T3 cells.

The structure and function of the epidermal growth factor receptor studied by using antisynthetic peptide antibodies

1985 ◽  
Vol 226 (1242) ◽  
pp. 127-134 ◽  
Keyword(s):  
Amino Acids ◽  
Epidermal Growth Factor Receptor ◽  
Amino Acid ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Amino Acid Sequence ◽  
Egf Receptor ◽  
Growth Factor Receptor ◽  
Binding Domain ◽  
Epidermal Growth

The human epidermal growth factor receptor has been purified and partial amino acid sequence obtained. A synthetic oligonucleotide was used to select complementary DNA clones from placental and A431 clone banks. The nucleotide sequence of a 5.8 kilobase transcript was determined and used to predict the total amino acid sequence of the receptor. We have predicted a model for the receptor which has an external ligand binding domain of 621 amino acids, a transmembrane region of 23 amino acids, and a cytoplasmic domain of 542 amino acids having protein tyrosine kinase activity. The kinase autophosphorylation sites have been mapped onto the primary amino acid sequence. Analysis of protein sequence databases have shown that the erb -B oncogene of avian erythroblastosis virus has acquired part of the avian EGF receptor gene. The hypothesis has been proposed that transformation by this virus is the result of expression of a truncated EGF receptor which lacks the majority of the EGF binding domain and delivers a continuous proliferation signal to transformed cells. We describe here the production of polyclonal and monoclonal antibodies to selected synthetic peptides from the EGF receptor and v-erb B sequences. Antisera to sequences encompassing the three major sites of autophosphorylation and the putative ATP binding site all recognize the native EGF receptor molecule. We have used these reagents to test our model of EGF receptor structure and v-erb B function.

scholarly journals A Single Amino Acid Exchange, Arg-45 to Ala, Generates an Epidermal Growth Factor (EGF) Mutant with High Affinity for the Chicken EGF Receptor

1995 ◽  
Vol 270 (38) ◽  
pp. 22337-22343 ◽  
Author(s):  
Monique L. M. van de Poll ◽  
Anne E. G. Lenferink ◽  
Marianne J. H. van Vugt ◽  
Jacqueline J. L. Jacobs ◽  
Jannie W. H. Janssen ◽  
...  
Keyword(s):  
Amino Acid ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Egf Receptor ◽  
Single Amino Acid ◽  
Amino Acid Exchange ◽  
High Affinity ◽  
Epidermal Growth ◽  
Acid Exchange

scholarly journals Nucleotide-binding properties of kinase-deficient epidermal-growth-factor- receptor mutants

1998 ◽  
Vol 330 (1) ◽  
pp. 353-359 ◽  
Author(s):  
Kunrong CHENG ◽  
G. John KOLAND
Keyword(s):  
Amino Acid ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Egf Receptor ◽  
Nucleotide Binding ◽  
Protein Kinase Activity ◽  
Amino Acid Substitutions ◽  
Wild Type ◽  
Binding Properties ◽  
Epidermal Growth

The nucleotide-binding properties of wild-type epidermal- growth-factor (EGF)-receptor protein tyrosine kinase (PTK) and EGF-receptor mutants with site-specific amino acid substitutions known to attenuate protein kinase activity were analysed by a fluorescence competition assay employing the nucleotide analogue 2ʹ(3ʹ)-O-(2,4,6-trinitrophenyl)adenosine 5ʹ-triphosphate.Binding affinities for ATP and Mn·ATP complex were determined for the PTK domains of the wild-type and two mutant proteins. Surprisingly, mutation of the highly conserved Lys-721 residue in the nucleotide-binding site of the EGF- receptor PTK domain did not abolish ATP and Mn·ATP binding, although the binding affinity for the Mn·ATP complex was significantly reduced. A second kinase-inactivating mutation that targeted the highly conserved Asp-813 residue had little effect on the nucleotide-binding properties of the EGF-receptor PTK domain. These results indicated that the principle effect of these two kinase-inactivating amino acid substitutions is not to block nucleotide binding, but is instead an inhibition of the phospho-transfer reaction.

947: Chemopreventive Effects of COX-2 Inhibitor and Epidermal Growth Factor (EGF)-Receptor Kinase Inhibitor on Rat Urinary Bladder Tumorigenesis

2004 ◽  
Vol 171 (4S) ◽  
pp. 251-251
Author(s):  
Kazunori Hattori ◽  
Katsuyuki Iida ◽  
Akira Johraku ◽  
Sadamu Tsukamoto ◽  
Taeko Asano ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Urinary Bladder ◽  
Egf Receptor ◽  
Kinase Inhibitor ◽  
Receptor Kinase ◽  
Rat Urinary Bladder ◽  
Epidermal Growth ◽  
Cox 2
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