The Epithelial Biochemistry Laboratory

Epithelial Biochemistry Laboratory

Structure-Function studies on the EGF/EGF receptor family

The EGF receptor family consists of four distinct tyrosine kinase receptors, EGFR/HER/ErbB1, HER2/Neu/ErbB2, HER3/ErbB3, and HER4/ErbB4 which interact with at least 12 different ligands which can induce homo- or heterodimerization of the family members. Much remains to be determined about the basis of ligand affinity and specificity to the receptors, the basis of homo- and heterodimerization and how this leads to activation of the intracellular tyrosine kinase domains of these ligands. We are currently persuing answers to some of these questions by expressing various receptor constructs and using a number of biophysical techniques to study them and their ligand interactions.

We have been part of a collaboration that recently solved the structure of TGF-alpha bound to an extracellular fragment of the EGF receptor that is able to bind ligand and dimerize. [More information]

Here, a biosensor was used to measure the binding of TGF-alpha and EGF on the G441K mutant of residues 1-501 of the EGF receptor. Note that the binding affinties of these ligands to the unmutated EGF receptor 1-501 are 35-40 and 13-21 nM, respectively. Thus, residue 441 of the EGF receptor is in the vicinity of the ligand binding site.

Lab members involved in the research:

Publications:
Crystal structure of a truncated epidermal growth factor receptor extracellular domain bound to transforming growth factor alpha
Garrett TPJ, McKern NM, Lou M, Elleman TC, Adams TE, Lovrecz GO, Zhu H-J, Walker F, Frenkel MJ, Hoyne PA, Jorissen RN, Nice EC, Burgess AW, Ward CW.
Cell. 110: 763-773 (2002). [Medline entry]

Identification of a determinant of epidermal growth factor receptor ligand-binding specificity using a truncated, high-affinity form of the ectodomain.
Elleman TC, Domagala T, McKern NM, Nerrie M, Lönnqvist B, Adams TE, Lewis J, Lovrecz GO, Hoyne PA, Richards KM, Howlett GJ, Rothacker J, Jorissen RN, Lou M, Garrett TPJ, Burgess AW, Nice EC, Ward CW.
Biochemistry 40:8930-8939 (2001). [Medline entry]

Stoichiometry, kinetic and binding analysis of the interaction between epidermal growth factor (EGF) and the extracellular domain of the EGF receptor.
Domagala T, Konstantopoulos N, Smyth F, Jorissen RN, Fabri L, Geleick D, Lax I, Schlessinger J, Sawyer W, Howlett GJ, Burgess AW, Nice EC.
Growth Factors 18:11-29 (2000). [Medline entry]

Structure-function relationships for the EGF/TGF-alpha family of mitogens.
Groenen LC, Nice EC, Burgess AW.
Growth Factors. 11:235-257 (1994). [Medline entry]

Solution structure of murine epidermal growth factor determined by NMR spectroscopy and refined by energy minimization with restraints.
Montelione GT, Wuthrich K, Burgess AW, Nice EC, Wagner G, Gibson KD, Scheraga HA.
Biochemistry. 31:236-249 (1992). [Medline entry]

Detection of colonic growth factors using a human colonic carcinoma cell line (LIM1215).
Whitehead RH, Nice EC, Lloyd CJ, James R, Burgess AW.
Int J Cancer. 46:858-863 (1990). [Medline entry]

Murine epidermal growth factor: structure and function.
Burgess AW, Lloyd CJ, Smith S, Stanley E, Walker F, Fabri L, Simpson RJ, Nice EC.
Biochemistry. Jul 12;27(14):4977-4985 (1988). [Medline entry]

Solution structure of murine epidermal growth factor: determination of the polypeptide backbone chain-fold by nuclear magnetic resonance and distance geometry.
Montelione GT, Wuthrich K, Nice EC, Burgess AW, Scheraga HA.
Proc Natl Acad Sci U S A. 84:5226-5230 (1987). [Medline entry]

Identification of two anti-parallel beta-sheet conformations in the solution structure of murine epidermal growth factor by proton magnetic resonance.
Montelione GT, Wuthrich K, Nice EC, Burgess AW, Scheraga HA.
Proc Natl Acad Sci U S A. 83:8594-8598 (1986). [Medline entry]

Binding of fluoresceinated epidermal growth factor to A431 cell sub-populations studied using a model-independent analysis of flow cytometric fluorescence data.
Chatelier RC, Ashcroft RG, Lloyd CJ, Nice EC, Whitehead RH, Sawyer WH, Burgess AW.
EMBO J. 5:1181-1186 (1986). [Medline entry]

Rat epidermal growth factor: complete amino acid sequence. Homology with the corresponding murine and human proteins; isolation of a form truncated at both ends with full in vitro biological activity.
Simpson RJ, Smith JA, Moritz RL, O'Hare MJ, Rudland PS, Morrison JR, Lloyd CJ, Grego B, Burgess AW, Nice EC.
Eur J Biochem. 153:629-637 (1985). [Medline entry]

A synthetic peptide containing the autophosphorylation site of the transforming protein of Harvey sarcoma virus is phosphorylated by the EGF-stimulated tyrosine kinase.
Baldwin GS, Stanley IJ, Nice EC.
FEBS Lett. 153:257-261 (1983). [Medline entry]

Murine epidermal growth factor: heterogeneity on high resolution ion-exchange chromatography.
Burgess AW, Lloyd CJ, Nice EC.
EMBO J. 2:2065-2069 (1983). [Medline entry]

Two forms of murine epidermal growth factor: rapid separation by using reverse-phase HPLC.
Burgess AW, Knesel J, Sparrow LG, Nicola NA, Nice EC.
Proc Natl Acad Sci U S A. 79:5753-5757 (1982). [Medline entry]