Abstract:
:The proteasome is a multi-subunit protein complex that serves as a major pathway for intracellular protein degradation, playing important functions in various biological processes. The C-terminus of the β7 (PSMB4) proteasome subunit was tagged with EGFP and with a composite element for affinity purification and TEV cleavage elution (HTBH). When the construct was retrovirally delivered into HeLa cells, virtually all of the β7-EGFP-HTBH fusion protein was found to be incorporated into fully functional proteasomes. This ensured that subcellular localization of the EGFP signal in living HeLa cells could be attributed to β7-EGFP-HTBH within the proteasome complex rather than to free protein. The β7-EGFP-HTBH fusion can, therefore, serve as a valuable tool for in vivo imaging of proteasomes as well as for high-affinity purification of these complexes and associated molecules for subsequent analyses.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Kulichkova VA,Artamonova TO,Zaykova JJ,Ermolaeva JB,Khodorkovskii MA,Barlev NA,Tomilin AN,Tsimokha ASdoi
10.1007/s12033-014-9799-0subject
Has Abstractpub_date
2015-01-01 00:00:00pages
36-44issue
1eissn
1073-6085issn
1559-0305journal_volume
57pub_type
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