Purification of active Escherichia coli ribosome recycling factor (RRF) from an osmo-regulated expression system.
Ribosome release factor (RRF) from Escherichia coli was overproduced from an osmo-expression vector. More than 40% of cell protein was RRF after 6 h of induction. A purification scheme is described that produced 50 mg of RRF from an initial culture of 2 L. The recycling time for ribosomes synthesising the tripeptide fMet-Phe-Leu in vitro in the absence of RF3 was reduced from 40 to 15 s by the addition of purified 1.5 microM RRF.
MacDougall
J
J
CNRS-UPR 9073, Institut de Biologie Physico-chimique, Paris, France.
Holst-Hansen
P
P
Mortensen
K K
KK
Freistroffer
D V
DV
Pavlov
M Y
MY
Ehrenberg
M
M
Buckingham
R H
RH
D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics; D011506:Proteins; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D012269:Ribosomal Proteins; D014882:Water-Electrolyte Balance
D001426:Bacterial Proteins; D011506:Proteins; D011994:Recombinant Proteins; D012269:Ribosomal Proteins; C087267:ribosome releasing factor