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Substrate screening identifies a novel target sequence for the proteasomal activity regulated by ionizing radiation
Author(s) -
BrogginiTenzer Angela,
Hollenstein Andreas,
Pianowski Zbigniew,
Wampfler Andrea,
Furmanova Polina,
Winssinger Nicolas,
Pruschy Martin
Publication year - 2010
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200900162
Subject(s) - proteasome , protease , cleavage (geology) , nucleoporin , ionizing radiation , biology , nuclear protein , microbiology and biotechnology , biochemistry , peptide sequence , nuclear pore , chemistry , computational biology , nuclear transport , enzyme , cell nucleus , nucleus , gene , transcription factor , irradiation , paleontology , physics , fracture (geology) , nuclear physics
The screening for treatment‐induced enzyme activities offers the opportunity to discover important regulatory mechanisms and the identification of potential targets for anti‐cancer therapies. A novel screening technique was applied to screen substrate peptide sequences for proteolytic activities up‐ or down‐regulated by ionizing radiation in tumor cells. One specific substrate sequence was cleaved in control cell extracts but to a smaller extent in irradiated cell extracts and investigated in detail. Based on protease‐class‐specific inhibitory studies and cleavage site analysis a potent warhead‐inhibitor was synthesized and used to identify the proteasome as the protease of interest. The investigated sequence shows high homology to a regulatory site of nucleoporin 50, an element of the nuclear pore complex, and site specific cleavage of nucleoporin 50 was determined in vitro suggesting a novel link between the ionizing radiation‐regulated proteasome and nuclear protein shuttling.