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DETERMINATION OF THE THREE‐DIMENSIONAL STRUCTURE OF FULL‐LENGTH HUMAN EPIDERMAL GROWTH FACTOR RECEPTOR BY CRYO‐ELECTRON TOMOGRAPHY
Author(s) -
Purba Endang R.,
Akhouri Reeresh R.,
Ofverstedt Larsgoran,
Skoglund Ulf,
Maruyama Ichiro
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.659.15
Subject(s) - autophosphorylation , epidermal growth factor receptor , epidermal growth factor , biophysics , transmembrane protein , extracellular , microbiology and biotechnology , ligand (biochemistry) , chemistry , receptor , biology , kinase , biochemistry , protein kinase a
Human epidermal growth factor receptor (EGFR) plays vital roles in cellular processes including cell proliferation, survival, motility and differentiation. Mice lacking EGFR die shortly after birth due to multi‐organ failure. Aberrant activation of EGFR is implicated in many human cancers. EGFR is synthesized as a single‐pass transmembrane protein, which consists of an extracellular ligand‐binding domain and an intracellular kinase domain separated by a single transmembrane domain. While crystal structures of EGFR domains are available, three‐dimensional structural determination of the full‐length EGFR has not been successful so far. In the conference, we will present cryo‐electron tomography structures of the intact human EGFR in the presence of bound EGF ligand. We successfully purified the full‐length EGFR with autophosphorylation activity upon EGF stimulation. We then prepared vitrified specimens and collected 30 tilt series projections spanning −60 to +60 degrees, with tilt increments of 0.5 degrees, using a FEI Titan Krios at 39,000× magnification and □1–2 μm underfocus. Tilt series data was analyzed with COMET software. These three‐dimensional structures of the full‐length EGFR are invaluable for understanding of mechanisms underlying activation of EGFR by ligand binding. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .