Abstract:
:Here we present a method to purify large amounts of highly pure and stably arrested ribosome-nascent chain complexes (RNCs) from Escherichia coli cells. It relies on the combined use of translation-arrest sequences to generate nascent polypeptides of specified length and subsequent tag purification of the RNCs. Moreover, we adapted this method for the in vivo production of RNCs with specific isotope labeling of the nascent chains for nuclear magnetic resonance (NMR) studies. This method opens therefore possibilities for a wide range of biochemical and structural studies exploring conformations of nascent chains during the early steps of protein folding and targeting.
journal_name
FEBS Lettjournal_title
FEBS lettersauthors
Rutkowska A,Beerbaum M,Rajagopalan N,Fiaux J,Schmieder P,Kramer G,Oschkinat H,Bukau Bdoi
10.1016/j.febslet.2009.06.041subject
Has Abstractpub_date
2009-07-21 00:00:00pages
2407-13issue
14eissn
0014-5793issn
1873-3468pii
S0014-5793(09)00499-2journal_volume
583pub_type
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