PMID: 19526984 RLIMS-P 3 eFIP 0 miRTex 0 eGARD 0 Issue Report
Title
| 1. | Characterizing the effects of the juxtamembrane domain on vascular endothelial growth factor receptor-2 enzymatic activity, autophosphorylation, and inhibition by axitinib. |
Abstract
| 2. | The catalytic domains of protein kinases are commonly treated as independent modular units with distinct biological functions. |
| 3. | Here, the interactions between the catalytic and juxtamembrane domains of VEGFR2 are studied. |
| 4. | Highly purified preparations of the receptor tyrosine kinase VEGFR2 catalytic domain without (VEGFR2-CD) and with (VEGFR2-CD/JM) the juxtamembrane (JM) domain were characterized by kinetic, biophysical, and structural methods. |
| 5. | Although the catalytic parameters for both constructs were similar, the autophosphorylation rate of VEGFR2-CD/JM was substantially faster than VEGFR2-CD. |
| 6. | The first event in the autophosphorylation reaction was phosphorylation of JM residue Y801 followed by phosphorylation of activation loop residues in the CD. |
| 7. | The rates of activation loop autophosphorylation for the two constructs were determined to be similar. |
| 8. | The autophosphorylation rate of Y801 was invariant on enzyme concentration, which is consistent with an intramolecular reaction. |
| 9. | In addition, the first biochemical characterization of the advanced clinical compound axitinib is reported. |
| 10. | Axitinib was found to have 40-fold enhanced biochemical potency toward VEGFR2-CD/JM (K(i) = 28 pM) compared to VEGFR2-CD, which correlates better with cellular potency. |
| 11. | Calorimetric studies, including a novel ITC compound displacement method, confirmed the potency and provided insight into the thermodynamic origin of the potency differences. |
| 12. | A structural model for the VEGFR2-CD/JM is proposed based on the experimental findings reported here and on the JM position in c-Kit, FLT3, and CSF1/cFMS. |
| 13. | The described studies identify potential functions of the VEGFR2 JM domain with implications to both receptor biology and inhibitor design. |
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Legend: SUBSTRATE KINASE INTERACTANT SITE GENE MIRNA ANOMALY EXPRESSION DISEASE OUTCOME/RESPONSE SR_DRUG DRUG CELL TRIGGER Normalized
Tool: RLIMS-P
| PTM enzyme | Substrate | Site | Sentence |
|---|---|---|---|
| VEGFR2-CD/JM (P35968) | VEGFR2-CD/JM (P35968) | 5 | |
| vascular endothelial growth factor receptor-2 enzymatic (P35968) | Tyr-801 | 1, 6 | |
| vascular endothelial growth factor receptor-2 enzymatic (P35968) | vascular endothelial growth factor receptor-2 enzymatic (P35968) | Tyr-801 | 1, 8 |