Visualizing GroEL/ES in the Act of Encapsulating a Folding Protein

Overview

Journal Cell

Publisher Cell Press

Specialty Cell Biology

Date 2013 Jun 11

PMID 23746846

Citations 61

Authors

Dong-Hua Chen

Damian Madan

Jeremy Weaver

Zong Lin

Gunnar F Schroder

Wah Chiu

Hays S Rye

Affiliations

Soon will be listed here.

Abstract

The GroEL/ES chaperonin system is required for the assisted folding of many proteins. How these substrate proteins are encapsulated within the GroEL-GroES cavity is poorly understood. Using symmetry-free, single-particle cryo-electron microscopy, we have characterized a chemically modified mutant of GroEL (EL43Py) that is trapped at a normally transient stage of substrate protein encapsulation. We show that the symmetric pattern of the GroEL subunits is broken as the GroEL cis-ring apical domains reorient to accommodate the simultaneous binding of GroES and an incompletely folded substrate protein (RuBisCO). The collapsed RuBisCO folding intermediate binds to the lower segment of two apical domains, as well as to the normally unstructured GroEL C-terminal tails. A comparative structural analysis suggests that the allosteric transitions leading to substrate protein release and folding involve concerted shifts of GroES and the GroEL apical domains and C-terminal tails.

Citing Articles

Exploring Novel Antibiotics by Targeting the GroEL/GroES Chaperonin System.

Wang Y, Tong Z, Han J, Li C, Chen X ACS Pharmacol Transl Sci. 2025; 8(1):10-20.

PMID: 39816798 PMC: 11729427. DOI: 10.1021/acsptsci.4c00397.

Visualizing chaperonin function in situ by cryo-electron tomography.

Wagner J, Carvajal A, Bracher A, Beck F, Wan W, Bohn S Nature. 2024; 633(8029):459-464.

PMID: 39169181 PMC: 11390479. DOI: 10.1038/s41586-024-07843-w.

Asymmetric apical domain states of mitochondrial Hsp60 coordinate substrate engagement and chaperonin assembly.

Braxton J, Shao H, Tse E, Gestwicki J, Southworth D Nat Struct Mol Biol. 2024; 31(12):1848-1858.

PMID: 38951622 PMC: 11638070. DOI: 10.1038/s41594-024-01352-0.

Structural basis of substrate progression through the bacterial chaperonin cycle.

Gardner S, Darrow M, Lukoyanova N, Thalassinos K, Saibil H Proc Natl Acad Sci U S A. 2023; 120(50):e2308933120.

PMID: 38064510 PMC: 10723157. DOI: 10.1073/pnas.2308933120.

Visualizing the chaperone-mediated folding trajectory of the G protein β5 β-propeller.

Wang S, Sass M, Kwon Y, Ludlam W, Smith T, Carter E Mol Cell. 2023; 83(21):3852-3868.e6.

PMID: 37852256 PMC: 10841713. DOI: 10.1016/j.molcel.2023.09.032.

References

Pettersen E, Goddard T, Huang C, Couch G, Greenblatt D, Meng E . UCSF Chimera--a visualization system for exploratory research and analysis. J Comput Chem. 2004; 25(13):1605-12. DOI: 10.1002/jcc.20084. View

Clare D, Bakkes P, Van Heerikhuizen H, Van Der Vies S, Saibil H . Chaperonin complex with a newly folded protein encapsulated in the folding chamber. Nature. 2009; 457(7225):107-10. PMC: 2728927. DOI: 10.1038/nature07479. View

Rye H, Roseman A, Chen S, Furtak K, Fenton W, Saibil H . GroEL-GroES cycling: ATP and nonnative polypeptide direct alternation of folding-active rings. Cell. 1999; 97(3):325-38. DOI: 10.1016/s0092-8674(00)80742-4. View

Cliff M, Limpkin C, Cameron A, Burston S, Clarke A . Elucidation of steps in the capture of a protein substrate for efficient encapsulation by GroE. J Biol Chem. 2006; 281(30):21266-21275. DOI: 10.1074/jbc.M601605200. View

Mayhew M, Da Silva A, Martin J, Erdjument-Bromage H, Tempst P, Hartl F . Protein folding in the central cavity of the GroEL-GroES chaperonin complex. Nature. 1996; 379(6564):420-6. DOI: 10.1038/379420a0. View

Fenton W, Kashi Y, Furtak K, Horwich A . Residues in chaperonin GroEL required for polypeptide binding and release. Nature. 1994; 371(6498):614-9. DOI: 10.1038/371614a0. View

Ueno T, Taguchi H, Tadakuma H, Yoshida M, Funatsu T . GroEL mediates protein folding with a two successive timer mechanism. Mol Cell. 2004; 14(4):423-34. DOI: 10.1016/s1097-2765(04)00261-8. View

Martin J, Langer T, Boteva R, Schramel A, Horwich A, Hartl F . Chaperonin-mediated protein folding at the surface of groEL through a 'molten globule'-like intermediate. Nature. 1991; 352(6330):36-42. DOI: 10.1038/352036a0. View

Elad N, Farr G, Clare D, Orlova E, Horwich A, Saibil H . Topologies of a substrate protein bound to the chaperonin GroEL. Mol Cell. 2007; 26(3):415-26. PMC: 1885994. DOI: 10.1016/j.molcel.2007.04.004. View

10.

Scheres S, Chen S . Prevention of overfitting in cryo-EM structure determination. Nat Methods. 2012; 9(9):853-4. PMC: 4912033. DOI: 10.1038/nmeth.2115. View

11.

Hartl F, Bracher A, Hayer-Hartl M . Molecular chaperones in protein folding and proteostasis. Nature. 2011; 475(7356):324-32. DOI: 10.1038/nature10317. View

12.

Baker M, Ju T, Chiu W . Identification of secondary structure elements in intermediate-resolution density maps. Structure. 2007; 15(1):7-19. PMC: 1810566. DOI: 10.1016/j.str.2006.11.008. View

13.

Kanno R, Koike-Takeshita A, Yokoyama K, Taguchi H, Mitsuoka K . Cryo-EM structure of the native GroEL-GroES complex from thermus thermophilus encapsulating substrate inside the cavity. Structure. 2009; 17(2):287-93. DOI: 10.1016/j.str.2008.12.012. View

14.

Clare D, Vasishtan D, Stagg S, Quispe J, Farr G, Topf M . ATP-triggered conformational changes delineate substrate-binding and -folding mechanics of the GroEL chaperonin. Cell. 2012; 149(1):113-23. PMC: 3326522. DOI: 10.1016/j.cell.2012.02.047. View

15.

Ludtke S, Baldwin P, Chiu W . EMAN: semiautomated software for high-resolution single-particle reconstructions. J Struct Biol. 1999; 128(1):82-97. DOI: 10.1006/jsbi.1999.4174. View

16.

Frank G, Goomanovsky M, Davidi A, Ziv G, Horovitz A, Haran G . Out-of-equilibrium conformational cycling of GroEL under saturating ATP concentrations. Proc Natl Acad Sci U S A. 2010; 107(14):6270-4. PMC: 2852005. DOI: 10.1073/pnas.0910246107. View

17.

Ranson N, Clare D, Farr G, Houldershaw D, Horwich A, Saibil H . Allosteric signaling of ATP hydrolysis in GroEL-GroES complexes. Nat Struct Mol Biol. 2006; 13(2):147-52. PMC: 2871290. DOI: 10.1038/nsmb1046. View

18.

Madan D, Lin Z, Rye H . Triggering protein folding within the GroEL-GroES complex. J Biol Chem. 2008; 283(46):32003-13. PMC: 2581556. DOI: 10.1074/jbc.M802898200. View

19.

Roseman A, Chen S, White H, Braig K, Saibil H . The chaperonin ATPase cycle: mechanism of allosteric switching and movements of substrate-binding domains in GroEL. Cell. 1996; 87(2):241-51. DOI: 10.1016/s0092-8674(00)81342-2. View

20.

Lin Z, Rye H . GroEL-mediated protein folding: making the impossible, possible. Crit Rev Biochem Mol Biol. 2006; 41(4):211-39. PMC: 3783267. DOI: 10.1080/10409230600760382. View