title: Triggering protein folding within the GroEL-GroES complex.
abstract: The folding of many proteins depends on the assistance of chaperonins like GroEL and GroES and involves the enclosure of substrate proteins inside an internal cavity that is formed when GroES binds to GroEL in the presence of ATP. Precisely how assembly of the GroEL-GroES complex leads to substrate protein encapsulation and folding remains poorly understood. Here we use a chemically modified mutant of GroEL (EL43Py) to uncouple substrate protein encapsulation from release and folding. Although EL43Py correctly initiates a substrate protein encapsulation reaction, this mutant stalls in an intermediate allosteric state of the GroEL ring, which is essential for both GroES binding and the forced unfolding of the substrate protein. This intermediate conformation of the GroEL ring possesses simultaneously high affinity for both GroES and non-native substrate protein, thus preventing escape of the substrate protein while GroES binding and substrate protein compaction takes place. Strikingly, assembly of the folding-active GroEL-GroES complex appears to involve a strategic delay in ATP hydrolysis that is coupled to disassembly of the old, ADP-bound GroEL-GroES complex on the opposite ring.
authors.affiliation: Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, USA.
authors.forename: Damian
authors.initials: D
authors.lastname: Madan
authors.forename: Zong
authors.initials: Z
authors.lastname: Lin
authors.forename: Hays S
authors.initials: HS
authors.lastname: Rye
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D018835:Chaperonin 10 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D018834:Chaperonin 60 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006868:Hydrolysis; D009154:Mutation / Q000235:genetics; D011485:Protein Binding; D017510:Protein Folding; D013379:Substrate Specificity
chemical_list: D018835:Chaperonin 10; D018834:Chaperonin 60; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases
references.citation: Mol Cell. 2007 May 11;26(3):415-26
references.pmid: 17499047
references.citation: J Mol Biol. 2004 May 14;338(5):969-77
references.pmid: 15111060
references.citation: J Mol Biol. 1994 Oct 28;243(3):397-401
references.pmid: 7966268
references.citation: Nat Struct Mol Biol. 2006 Feb;13(2):147-52
references.pmid: 16429154
references.citation: J Biol Chem. 2003 Jun 27;278(26):23311-6
references.pmid: 12660239
references.citation: Cell. 1999 Apr 30;97(3):325-38
references.pmid: 10319813
references.citation: J Biol Chem. 2006 Jul 28;281(30):21266-21275
references.pmid: 16684774
references.citation: Methods. 2001 Jul;24(3):278-88
references.pmid: 11403576
references.citation: Proc Natl Acad Sci U S A. 2004 May 11;101(19):7264-9
references.pmid: 15123793
references.citation: Nature. 1997 Aug 21;388(6644):792-8
references.pmid: 9285593
references.citation: Nature. 1994 Oct 13;371(6498):614-9
references.pmid: 7935796
references.citation: Mol Cell. 2004 May 21;14(4):423-34
references.pmid: 15149592
references.citation: Nature. 1991 Jul 4;352(6330):36-42
references.pmid: 1676490
references.citation: Proc Natl Acad Sci U S A. 2004 Aug 24;101(34):12461-6
references.pmid: 15314233
references.citation: Cell. 1996 Oct 18;87(2):241-51
references.pmid: 8861908
references.citation: Crit Rev Biochem Mol Biol. 2006 Jul-Aug;41(4):211-39
references.pmid: 16849107
references.citation: Methods Enzymol. 1983;100:144-60
references.pmid: 6312256
references.citation: Q Rev Biophys. 2003 May;36(2):229-56
references.pmid: 14686103
references.citation: Structure. 1996 Jan 15;4(1):79-88
references.pmid: 8805508
references.citation: Biochemistry. 1991 Feb 12;30(6):1586-91
references.pmid: 1671555
references.citation: Proc Natl Acad Sci U S A. 2000 Aug 15;97(17):9367-72
references.pmid: 10944209
references.citation: Biochemistry. 1991 Sep 24;30(38):9195-200
references.pmid: 1680001
references.citation: Nature. 1994 Oct 13;371(6498):578-86
references.pmid: 7935790
references.citation: Cell. 1994 Aug 26;78(4):693-702
references.pmid: 7915201
references.citation: J Biol Chem. 2008 Mar 14;283(11):6886-96
references.pmid: 18184659
references.citation: Science. 1994 Jul 29;265(5172):659-66
references.pmid: 7913555
references.citation: Proc Natl Acad Sci U S A. 2007 Mar 27;104(13):5342-7
references.pmid: 17372195
references.citation: Cell. 1996 Feb 9;84(3):481-90
references.pmid: 8608602
references.citation: Cell. 1995 Nov 17;83(4):577-87
references.pmid: 7585961
references.citation: J Mol Biol. 1995 May 26;249(1):138-52
references.pmid: 7776368
references.citation: Nature. 1989 Dec 21-28;342(6252):884-9
references.pmid: 10532860
references.citation: Biochemistry. 1991 Oct 8;30(40):9716-23
references.pmid: 1680394
references.citation: Nat Struct Mol Biol. 2008 Mar;15(3):303-11
references.pmid: 18311152
references.citation: J Mol Biol. 1998 Apr 24;278(1):267-78
references.pmid: 9571049
references.citation: J Biol Chem. 1996 Nov 8;271(45):28229-34
references.pmid: 8910440
references.citation: Nature. 1994 Sep 15;371(6494):261-4
references.pmid: 7915827
references.citation: Nat Rev Mol Cell Biol. 2004 Oct;5(10):781-91
references.pmid: 15459659
references.citation: Biochemistry. 1995 Apr 25;34(16):5303-8
references.pmid: 7727391
references.citation: J Biol Chem. 1997 May 30;272(22):14080-6
references.pmid: 9162032
references.citation: Cell. 2001 Dec 28;107(7):869-79
references.pmid: 11779463
references.citation: Cell. 1999 Dec 23;99(7):757-68
references.pmid: 10619429
references.citation: Nature. 1997 Aug 21;388(6644):741-50
references.pmid: 9285585
references.citation: Mol Cell. 2004 Oct 8;16(1):23-34
references.pmid: 15469819
references.citation: Cell. 2006 Jun 2;125(5):903-14
references.pmid: 16751100
references.citation: Proc Natl Acad Sci U S A. 2006 May 16;103(20):7595-600
references.pmid: 16682620
references.citation: Nature. 1996 Feb 1;379(6564):420-6
references.pmid: 8559246