Large-scale purification of ribosome-nascent chain complexes for biochemical and structural studies.

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 Lett

journal_title

FEBS letters

authors

Rutkowska A,Beerbaum M,Rajagopalan N,Fiaux J,Schmieder P,Kramer G,Oschkinat H,Bukau B

doi

10.1016/j.febslet.2009.06.041

subject

Has Abstract

pub_date

2009-07-21 00:00:00

pages

2407-13

issue

14

eissn

0014-5793

issn

1873-3468

pii

S0014-5793(09)00499-2

journal_volume

583

pub_type

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