Determination of the number of active GroES subunits in the fused heptamer GroES required for interactions with GroEL

The Journal of Biological Chemistry
Tatsuya NojimaFumihiro Motojima

Abstract

A double-heptamer ring chaperonin GroEL binds denatured substrate protein, ATP, and GroES to the same heptamer ring and encapsulates substrate into the central cavity underneath GroES where productive folding occurs. GroES is a disk-shaped heptamer, and each subunit has a GroEL-binding loop. The residues of the GroEL subunit responsible for GroES binding largely overlap those involved in substrate binding, and the mechanism by which GroES can replace the substrate when GroES binds to GroEL/substrate complex remains to be clarified. To address this question, we generated single polypeptide GroES by fusing seven subunits with various combinations of active and GroEL binding-defective subunits. Functional tests of the fused GroES variants indicated that four active GroES subunits were required for efficient formation of the stable GroEL/GroES complex and five subunits were required for the productive GroEL/substrate/GroES complex. An increase in the number of defective GroES subunits resulted in a slowing of encapsulation and folding. These results indicate the presence of an intermediate GroEL/substrate/GroES complex in which the substrate and GroES bind to GroEL by sharing seven common binding sites.

References

Mar 1, 1992·Protein Science : a Publication of the Protein Society·P V ViitanenG H Lorimer
May 26, 1995·Journal of Molecular Biology·S G BurstonA R Clarke
May 9, 1997·The Journal of Biological Chemistry·Y MakinoM Yoshida
Feb 26, 1998·Cell·B Bukau, A L Horwich
Oct 6, 1998·Annual Review of Biochemistry·P B SiglerA L Horwich
Jul 20, 1999·The Journal of Biological Chemistry·C SakikawaM Yoshida
Jan 26, 2000·Nature·W A HouryF U Hartl
Feb 16, 2000·Biochemical and Biophysical Research Communications·F MotojimaM Yoshida
May 8, 2001·Annual Review of Biophysics and Biomolecular Structure·D Thirumalai, G H Lorimer
Dec 7, 2002·Trends in Biochemical Sciences·Helen R Saibil, Neil A Ranson
May 9, 2003·The Journal of Biological Chemistry·Fumihiro Motojima, Masasuke Yoshida
May 20, 2004·Molecular Cell·Taro UenoTakashi Funatsu
Oct 22, 2005·The Journal of Biological Chemistry·Ayumi Koike-TakeshitaHideki Taguchi
May 15, 2007·Molecular Cell·Nadav EladHelen R Saibil

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Citations

Jul 30, 2009·Quarterly Reviews of Biophysics·Arthur L Horwich, Wayne A Fenton
Sep 13, 2013·Nature Reviews. Molecular Cell Biology·Helen Saibil
Jun 1, 2010·The Journal of Biological Chemistry·Tomoya SameshimaTakashi Funatsu
Jul 16, 2011·The Journal of Biological Chemistry·Melissa IllingworthLingling Chen
Apr 2, 2010·Cellular and Molecular Life Sciences : CMLS·Raimon SabateSalvador Ventura
Oct 21, 2010·The EMBO Journal·Fumihiro Motojima, Masasuke Yoshida
May 29, 2012·Journal of Molecular Biology·Tatsuya NojimaMasasuke Yoshida
Jul 24, 2013·Biopolymers·Daniel K Clare, Helen R Saibil
Feb 20, 2020·Quarterly Reviews of Biophysics·Arthur L Horwich, Wayne A Fenton
Jan 1, 2015·Biophysics·Fumihiro Motojima

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