Crystallization of the first three domains of the human insulin-like growth factor-1 receptor

Protein Sci. 1997 Dec;6(12):2663-6. doi: 10.1002/pro.5560061223.

Abstract

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 A resolution with cell dimensions a = 77.0 A, b = 99.5 A, c = 120.1 A, and space group P2(1)2(1)2(1).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • CHO Cells
  • Chromatography, Gel
  • Chromatography, Ion Exchange
  • Cricetinae
  • Crystallization
  • Crystallography, X-Ray
  • Humans
  • Molecular Sequence Data
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / isolation & purification
  • Receptor, IGF Type 1 / chemistry*
  • Receptor, IGF Type 1 / genetics
  • Recombinant Proteins
  • Transfection

Substances

  • Peptide Fragments
  • Recombinant Proteins
  • Receptor, IGF Type 1