GroEL/GroES cycling: ATP binds to an open ring before substrate protein favoring protein binding and production of the native state.

Proceedings of the National Academy of Sciences of the United States of America
Navneet K TyagiA L Horwich

Abstract

The GroEL/GroES reaction cycle involves steps of ATP and polypeptide binding to an open GroEL ring before the GroES encapsulation step that triggers productive folding in a sequestered chamber. The physiological order of addition of ATP and nonnative polypeptide, typically to the open trans ring of an asymmetrical GroEL/GroES/ADP complex, has been unknown, although there have been assumptions that polypeptide binds first, allowing subsequent ATP-mediated movement of the GroEL apical domains to exert an action of forceful unfolding on the nonnative polypeptide. Here, using fluorescence measurements, we show that the physiological order of addition is the opposite, involving rapid binding of ATP, accompanied by nearly as rapid apical domain movements, followed by slower binding of nonnative polypeptide. In order-of-addition experiments, approximately twice as much Rubisco activity was recovered when nonnative substrate protein was added after ATP compared with it being added before ATP, associated with twice as much Rubisco protein recovered with the chaperonin. Furthermore, the rate of Rubisco binding to an ATP-exposed ring was twice that observed in the absence of nucleotide. Finally, when both ATP and Rubisco were added simult...Continue Reading

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Citations

Mar 27, 2013·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Roy A Quinlan, R John Ellis
Mar 29, 2011·Cold Spring Harbor Perspectives in Biology·Axel MogkBernd Bukau
Nov 10, 2013·PloS One·Toshifumi MizutaTomohiro Mizobata
Oct 30, 2013·Proceedings of the National Academy of Sciences of the United States of America·Xiang Ye, George H Lorimer
Aug 20, 2014·Proceedings of the National Academy of Sciences of the United States of America·Xue FeiGeorge H Lorimer
Jul 5, 2015·FEBS Letters·Lars SkjærvenJosé María Valpuesta
May 31, 2011·Journal of Molecular Biology·Jin ChenKunihiro Kuwajima
Dec 18, 2014·Biophysical Journal·Hagen HofmannBenjamin Schuler
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Apr 22, 2017·The Journal of Physical Chemistry. B·Jin ChenWenfei Li
Aug 4, 2018·Journal of Medical Microbiology·Ciamak Ghazaei, Marie Line El Helou
Jan 30, 2016·Science Advances·Ching-Wen HoGiovanni Maglia
Jul 24, 2013·Biopolymers·Daniel K Clare, Helen R Saibil

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