EMBO J 2008,
PMID: 18273061
Ng, Cherlyn; Jackson, Rebecca A; Buschdorf, Jan P; Sun, Qingxiang; Guy, Graeme R; Sivaraman, J
The c-Cbl tyrosine kinase binding domain (Cbl-TKB), essentially an 'embedded' SH2 domain, has a critical role in targeting proteins for ubiquitination. To address how this domain can bind to disparate recognition mofits and to determine whether this results in variations in substrate-binding affinity, we compared crystal structures of the Cbl-TKB domain complexed with phosphorylated peptides of Sprouty2, Sprouty4, epidermal growth factor receptor, Syk, and c-Met receptors and validated the binding with point-mutational analyses using full-length proteins. An obligatory, intrapeptidyl H-bond between the phosphotyrosine and the conserved asparagine or adjacent arginine is essential for binding and orients the peptide into a positively charged pocket on c-Cbl. Surprisingly, c-Met bound to Cbl in the reverse direction, which is unprecedented for SH2 domain binding. The necessity of this intrapeptidyl H-bond was confirmed with isothermal titration calorimetry experiments that also showed Sprouty2 to have the highest binding affinity to c-Cbl; this may enable the selective sequestration of c-Cbl from other target proteins.
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Text Mining Data
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Manually curated Databases
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IRef Biogrid Interaction:
CBL
—
SPRY2
(physical association, affinity chromatography technology)
-
IRef Biogrid Interaction:
CBL
—
SPRY2
(association, x-ray crystallography)
-
IRef Biogrid Interaction:
CBL
—
GRB2
(physical association, affinity chromatography technology)
-
IRef Biogrid Interaction:
CBL
—
MET
(association, x-ray crystallography)
-
IRef Biogrid Interaction:
CBL
—
MET
(physical association, affinity chromatography technology)
-
IRef Biogrid Interaction:
CBL
—
EGFR
(association, x-ray crystallography)
-
IRef Biogrid Interaction:
CBL
—
EGFR
(physical association, affinity chromatography technology)
-
IRef Biogrid Interaction:
SH2B2
—
CBL
(physical association, affinity chromatography technology)
-
IRef Biogrid Interaction:
CBL
—
SPRY4
(physical association, affinity chromatography technology)
-
IRef Hprd Interaction:
SYK
—
CBL
(in vitro)
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IRef Hprd Interaction:
SYK
—
CBL
(in vivo)
-
IRef Hprd Interaction:
CBL
—
SPRY2
(in vivo)
-
IRef Hprd Interaction:
CBL
—
SPRY2
(in vitro)
-
IRef Hprd Interaction:
CBL
—
SPRY2
(two hybrid)
-
IRef Hprd Interaction:
CBL
—
GRB2
(in vitro)
-
IRef Hprd Interaction:
CBL
—
GRB2
(in vivo)
-
IRef Hprd Interaction:
CBL
—
MET
(in vitro)
-
IRef Hprd Interaction:
CBL
—
MET
(two hybrid)
-
IRef Hprd Interaction:
CBL
—
MET
(in vivo)
-
IRef Hprd Interaction:
CBL
—
EGFR
(in vivo)
-
IRef Hprd Interaction:
CBL
—
EGFR
(in vitro)
-
IRef Hprd Interaction:
SRC
—
CBL
(in vivo)
-
IRef Hprd Interaction:
SRC
—
CBL
(in vitro)
-
IRef Hprd Interaction:
FNBP1
—
CBL
(in vitro)
-
IRef Hprd Interaction:
FNBP1
—
CBL
(in vivo)
-
IRef Hprd Interaction:
SH2B2
—
CBL
(in vivo)
-
IRef Hprd Interaction:
SH2B2
—
CBL
(in vitro)
-
IRef Intact Interaction:
SYK
—
CBL
(direct interaction, isothermal titration calorimetry)
-
IRef Intact Interaction:
SYK
—
CBL
(direct interaction, x-ray crystallography)
-
IRef Intact Interaction:
CBL
—
SPRY2
(direct interaction, x-ray crystallography)
-
IRef Intact Interaction:
CBL
—
SPRY2
(direct interaction, isothermal titration calorimetry)
-
IRef Intact Interaction:
CBL
—
SPRY2
(physical association, anti tag coimmunoprecipitation)
-
IRef Intact Interaction:
GRB2
—
CBL
(physical association, anti bait coimmunoprecipitation)
-
IRef Intact Interaction:
CBL
—
MST1R
(direct interaction, isothermal titration calorimetry)
-
IRef Intact Interaction:
CBL
—
MET
(direct interaction, isothermal titration calorimetry)
-
IRef Intact Interaction:
CBL
—
MET
(physical association, anti bait coimmunoprecipitation)
-
IRef Intact Interaction:
CBL
—
MET
(direct interaction, x-ray crystallography)
-
IRef Intact Interaction:
CBL
—
EGFR
(physical association, anti bait coimmunoprecipitation)
-
IRef Intact Interaction:
CBL
—
EGFR
(direct interaction, x-ray crystallography)
-
IRef Intact Interaction:
CBL
—
EGFR
(direct interaction, isothermal titration calorimetry)
-
IRef Intact Interaction:
SRC
—
CBL
(direct interaction, isothermal titration calorimetry)
-
IRef Intact Interaction:
FLT1
—
CBL
(direct interaction, isothermal titration calorimetry)
-
IRef Intact Interaction:
SH2B2
—
CBL
(direct interaction, isothermal titration calorimetry)
-
IRef Intact Interaction:
SH2B2
—
CBL
(physical association, anti tag coimmunoprecipitation)
-
IRef Intact Interaction:
SPRY4
—
CBL
(physical association, anti tag coimmunoprecipitation)
-
IRef Intact Interaction:
SPRY4
—
CBL
(direct interaction, x-ray crystallography)
-
IRef Intact Interaction:
SPRY4
—
CBL
(direct interaction, isothermal titration calorimetry)
In total, 11 gene pairs are associated to this article in curated databases