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The same site on the integrase-binding domain of lens epithelium-derived growth factor is a therapeutic target for MLL leukemia and HIV.

Blood. 2014; 
Murai Marcelo J,Pollock Jonathan,He Shihan,Miao Hongzhi,Purohit Trupta,Yokom Adam,Hess Jay L,Muntean Andrew G,Grembecka Jolanta,Cierpicki To
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Bacterial Expression System Methods Molecular biology LEDGF constructs for bacterial expression have been ordered from GenScript and cloned into pET32a.... Forty-eight hours after transfection, whole-cells lysates were immunoprecipitated with anti-FLAG M-2 Magnetic Beads (M8823) (Sigma-Aldrich) and analyzed by sodium dodecyl sulfate– polyacrylamide gel electrophoresis and western blotting using anti-N-MLL antibody (05-764; Millipore), anti-b-actin antibody (A00702; GenScript), anti-menin antibody, and anti-HA tag antibody (2367S; Cell Signaling). Get A Quote

摘要

Lens epithelium-derived growth factor (LEDGF) is a chromatin-associated protein implicated in leukemia and HIV type 1 infection. LEDGF associates with mixed-lineage leukemia (MLL) fusion proteins and menin and is required for leukemic transformation. To better understand the molecular mechanism underlying the LEDGF integrase-binding domain (IBD) interaction with MLL fusion proteins in leukemia, we determined the solution structure of the MLL-IBD complex. We found a novel MLL motif, integrase domain binding motif 2 (IBM2), which binds to a well-defined site on IBD. Point mutations within IBM2 abolished leukemogenic transformation by MLL-AF9, validating that this newly identified motif is essential for th... More

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