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Crystallization of the first three domains of the human insulin‐like growth factor‐1 receptor
Author(s) -
Mckern Neil M.,
Frenkel Maurice J.,
Verkuylen Amanda,
Bentley John D.,
Lovrecz George O.,
Ivancic Neva,
Elleman Thomas C.,
Cosgrove Leah J.,
Lou Meizhen,
Garrett THOMAS P. J.,
Ward Colin W.
Publication year - 1997
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.5560061223
Subject(s) - ectodomain , monoclonal antibody , receptor , tyrosine kinase , growth factor , chemistry , size exclusion chromatography , insulin receptor , insulin like growth factor , antibody , biochemistry , microbiology and biotechnology , biology , insulin , endocrinology , insulin resistance , immunology , enzyme
The insulin‐like growth factor‐1 receptor (IGF‐1R) is a tyrosine kinase receptor of central importance in cell proliferation. A fragment (residues 1‐462) comprising the L1‐cysteine rich‐L2 domains of the human IGF‐1R ectodomain has been overexpressed in glycosylation‐deficient Lec8 cells and has been affinity‐purified via a c‐myc tag followed by gel filtration. The fragment was recognized by two anti‐IGF‐1R monoclonal antibodies, 24‐31 and 24‐60, but showed no detectable binding of IGF‐1 or IGF‐2. Isocratic elution of IGF‐1R/462 on anion‐exchange chromatography reduced sample heterogeneity, permitting the production of crystals that diffracted to 2.6 Å resolution with cell dimensions a = 77.0 A, Å = 99.5 Å, c = 120.1 Å, and space group P 2 1 2 1 2 1 .