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Characterization and inhibition of AF10‐mediated interaction
Author(s) -
Hagen Sven,
Mattay Dinah,
Räuber Christina,
Müller Kristian M.,
Arndt Katja M.
Publication year - 2014
Publication title -
journal of peptide science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 66
eISSN - 1099-1387
pISSN - 1075-2617
DOI - 10.1002/psc.2626
Subject(s) - coiled coil , peptide , phage display , fusion protein , biology , gene , computational biology , k562 cells , chromosomal translocation , microbiology and biotechnology , chemistry , biochemistry , recombinant dna
The non‐random chromosomal translocations t(10;11)(p13;q23) and t(10;11)(p13;q14–21) result in leukemogenic fusion proteins comprising the coiled coil domain of the transcription factor AF10 and the proteins MLL or CALM, respectively, and subsequently cause certain types of acute leukemia. The AF10 coiled‐coil domain, which is crucial for the leukemogenic effect, has been shown to interact with GAS41, a protein previously identified as the product of an amplified gene in glioblastoma. Using sequential synthetic peptides, we mapped the potential AF10/GAS41 interaction site, which was subsequently be used as scaffold for a library targeting the AF10 coiled‐coil domain. Using phage display, we selected a peptide that binds the AF10 coiled‐coil domain with higher affinity than the respective coiled‐coil region of wild‐type GAS41, as demonstrated by phage ELISA, CD, and PCAs. Furthermore, we were able to successfully deploy the inhibitory peptide in a mammalian cell line to lower the expression of Hoxa genes that have been described to be overexpressed in these leukemias. This work dissects molecular determinants mediating AF10‐directed interactions in leukemic fusions comprising the N‐terminal parts of the proteins MLL or CALM and the C‐terminal coiled‐coil domain of AF10. Furthermore, it outlines the first steps in recognizing and blocking the leukemia‐associated AF10 interaction in histiocytic lymphoma cells and therefore, may have significant implications in future diagnostics and therapeutics. Copyright © 2014 European Peptide Society and John Wiley & Sons, Ltd.