Crystallization of the first three domains of the human insulin-like growth factor-1 receptor.
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).
McKern
N M
NM
CSIRO Division of Molecular Science, Parkville, Victoria, Australia.
Lou
M
M
Frenkel
M J
MJ
Verkuylen
A
A
Bentley
J D
JD
Lovrecz
G O
GO
Ivancic
N
N
Elleman
T C
TC
Garrett
T P
TP
Cosgrove
L J
LJ
Ward
C W
CW
D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D006224:Cricetinae; D003460:Crystallization; D018360:Crystallography, X-Ray; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D017526:Receptor, IGF Type 1 / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins; D014162:Transfection
D010446:Peptide Fragments; D011994:Recombinant Proteins; D017526:Receptor, IGF Type 1