The chaperonin GroEL is destabilized by binding of ADP.


Abstract

The urea-induced dissociation and subsequent conformational transitions of the nucleotide-bound form of GroEL were studied by light scattering, 4,4'-bis(1-anilino-8- naphthalenesulfonic acid) binding, and intrinsic tyrosine fluorescence. Magnesium ion alone (10 mM) stabilizes GroEL and leads to coordination of the structural transitions monitored by the different parameters. The midpoint of the light-scattering transition that monitored dissociation of the 14-mer with bound magnesium was raised to approximately 3 M, which is considerably higher than the ligand-free form of the protein, which exhibits a transition with a midpoint at approximately 2 M urea. Binding of ADP results in destabilization of the GroEL oligomeric structure, and complete dissociation of the 14-mer in the presence of 5 mM ADP occurs at about 2 M urea with the midpoint of the transition at approximately 1 M urea. The same destabilization by ADP and stabilization by Mg2+ were seen when the conformation was followed by the intrinsic fluorescence. Complexation with the nonhydrolyzable ATP analog, 5'-adenylimidodiphosphate gave an apparent stability of the quaternary structure that was between that observed with Mg2+ and that with ADP. The ADP-bound form of the protein demonstrated increased hydrophobic exposure at lower urea concentrations than the uncomplexed GroEL. In addition, the GroEL-ADP complex is more accessible for proteolytic digestion by chymotrypsin than the uncomplexed protein, consistent with a more open, flexible form of the protein. The implication of the conformational changes to the mechanism of the GroEL function is discussed. Study holds ProTherm entries: 5205, 5206 Extra Details:

Submission Details

ID: cknCa8o33

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:29 p.m.

Version: 1

Publication Details
Gorovits BM;Horowitz PM,J. Biol. Chem. (1995) The chaperonin GroEL is destabilized by binding of ADP. PMID:7499369
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
3VZ6 2013-11-13 1.5 Crystal Structure Analysis of the Mini-chaperonines, variant with Gly 184 replaced with Ile and Leu 185 replaced Val and Val 186 replaced with Leu.
1KID 1997-09-17 1.7 GROEL (HSP60 CLASS) FRAGMENT (APICAL DOMAIN) COMPRISING RESIDUES 191-376, MUTANT WITH ALA 262 REPLACED WITH LEU AND ILE 267 REPLACED WITH MET
3VZ7 2013-11-13 1.8 Crystal Structure Analysis of the mini-chaperonin variant with Pro 187 Gly
3VZ8 2013-11-13 1.9 Crystal Structure Analysis of the Mini-chaperonin variant with Leu 185, Val 186, Pro 187, Arg 188 and Ser 190 replaced with all Gly
1GRU 2002-01-28 12.5 SOLUTION STRUCTURE OF GROES-ADP7-GROEL-ATP7 COMPLEX BY CRYO-EM
2C7E 2006-02-16 14.9 REVISED ATOMIC STRUCTURE FITTING INTO A GROEL(D398A)-ATP7 CRYO-EM MAP (EMD 1047)
3ZQ1 2013-06-19 15.9 Visualizing GroEL-ES in the Act of Encapsulating a Non-Native Substrate Protein
1SX3 2005-03-01 2.0 GroEL14-(ATPgammaS)14
1KP8 2003-03-25 2.0 Structural Basis for GroEL-assisted Protein Folding from the Crystal Structure of (GroEL-KMgATP)14 at 2.0 A Resolution
1DK7 2000-01-05 2.02 CRYSTAL STRUCTURE OF AN ISOLATED APICAL DOMAIN OF GROEL
1LA1 2002-04-03 2.06 Gro-EL Fragment (Apical Domain) Comprising Residues 188-379
1DKD 2000-01-12 2.1 CRYSTAL STRUCTURE OF A GROEL (APICAL DOMAIN) AND A DODECAMERIC PEPTIDE COMPLEX
1FYA 2000-11-22 2.2 CRYSTAL STRUCTURE OF THE HEXA-SUBSTITUTED MUTANT OF THE MOLECULAR CHAPERONIN GROEL APICAL DOMAIN
1FY9 2000-11-22 2.2 CRYSTAL STRUCTURE OF THE HEXA-SUBSTITUTED MUTANT OF THE MOLECULAR CHAPERONIN GROEL APICAL DOMAIN
1JON 1997-03-12 2.5 GROEL (HSP60 CLASS) FRAGMENT COMPRISING RESIDUES 191-345
1SS8 2005-03-01 2.7 GroEL
1OEL 1996-04-03 2.8 CONFORMATIONAL VARIABILITY IN THE REFINED STRUCTURE OF THE CHAPERONIN GROEL AT 2.8 ANGSTROM RESOLUTION
1GRL 1995-10-15 2.8 THE CRYSTAL STRUCTURE OF THE BACTERIAL CHAPERONIN GROEL AT 2.8 ANGSTROMS
1SVT 2005-03-01 2.81 Crystal structure of GroEL14-GroES7-(ADP-AlFx)7
1PCQ 2003-10-14 2.81 Crystal structure of groEL-groES
1XCK 2005-10-25 2.92 Crystal structure of apo GroEL
1PF9 2003-11-04 2.99 GroEL-GroES-ADP
1SX4 2005-03-01 3.0 GroEL-GroES-ADP7
1MNF 2003-10-07 3.0 Domain motions in GroEL upon binding of an oligopeptide
1AON 1997-10-15 3.0 CRYSTAL STRUCTURE OF THE ASYMMETRIC CHAPERONIN COMPLEX GROEL/GROES/(ADP)7
2NWC 2007-05-22 3.02 A 3.02 angstrom crystal structure of wild-type apo GroEL in a monoclinic space group
4WSC 2015-11-11 3.04 Crystal structure of a GroELK105A mutant
4WGL 2015-09-30 3.13 Crystal structure of a GroEL D83A/R197A double mutant
5OPW 2018-01-10 3.19 Crystal structure of the GroEL mutant A109C
2EU1 2006-08-29 3.29 Crystal structure of the chaperonin GroEL-E461K
5W0S 2017-08-09 3.5 GroEL using cryoEM
4V43 2014-07-09 3.52 Structural and mechanistic basis for allostery in the bacterial chaperonin GroEL
5OPX 2018-01-10 3.64 Crystal structure of the GroEL mutant A109C in complex with GroES and ADP BeF2
3WVL 2014-09-17 3.79 Crystal structure of the football-shaped GroEL-GroES complex (GroEL: GroES2:ATP14) from Escherichia coli
3E76 2009-08-25 3.94 Crystal structure of Wild-type GroEL with bound Thallium ions
3CAU 2008-09-02 4.2 D7 symmetrized structure of unliganded GroEL at 4.2 Angstrom resolution by cryoEM
2YEY 2011-05-18 4.5 Crystal structure of the allosteric-defective chaperonin GroEL E434K mutant
3C9V 2008-09-02 4.7 C7 Symmetrized Structure of Unliganded GroEL at 4.7 Angstrom Resolution from CryoEM
2C7C 2006-01-25 7.7 FITTED COORDINATES FOR GROEL-ATP7-GROES CRYO-EM COMPLEX (EMD-1180)
1GR5 2002-01-28 7.9 Solution Structure of apo GroEL by Cryo-Electron microscopy
4AAQ 2012-12-12 8.0 ATP-triggered molecular mechanics of the chaperonin GroEL
4AAR 2012-12-12 8.0 ATP-triggered molecular mechanics of the chaperonin GroEL
2CGT 2006-03-29 8.2 GROEL-ADP-gp31 COMPLEX
2YNJ 2012-12-12 8.4 GroEL at sub-nanometer resolution by Constrained Single Particle Tomography
4AB3 2012-12-12 8.5 ATP-triggered molecular mechanics of the chaperonin GroEL
4AB2 2012-12-12 8.5 ATP-triggered molecular mechanics of the chaperonin GroEL
4AAU 2012-12-12 8.5 ATP-triggered molecular mechanics of the chaperonin GroEL
4AAS 2012-12-12 8.5 ATP-triggered molecular mechanics of the chaperonin GroEL
2C7D 2006-01-25 8.7 Fitted coordinates for GroEL-ADP7-GroES Cryo-EM complex (EMD-1181)
3ZPZ 2013-06-19 8.9 Visualizing GroEL-ES in the Act of Encapsulating a Non-Native Substrate Protein
3ZQ0 2013-06-19 9.2 Visualizing GroEL-ES in the Act of Encapsulating a Non-Native Substrate Protein

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
90.2 60 kDa chaperonin Q8KIX2 CH60_BUCTS
91.7 60 kDa chaperonin P0C193 CH60_SODGL
91.3 60 kDa chaperonin A1JIP3 CH60_YERE8
91.9 60 kDa chaperonin Q2NW94 CH60_SODGM
91.5 60 kDa chaperonin P48219 CH60_YEREN
91.7 60 kDa chaperonin B1JMR1 CH60_YERPY
91.7 60 kDa chaperonin Q66FD5 CH60_YERPS
91.7 60 kDa chaperonin A4TRR0 CH60_YERPP
91.7 60 kDa chaperonin Q1CED4 CH60_YERPN
91.7 60 kDa chaperonin A9QYQ1 CH60_YERPG
91.7 60 kDa chaperonin Q8ZIY3 CH60_YERPE
91.7 60 kDa chaperonin B2K1Y4 CH60_YERPB
91.7 60 kDa chaperonin Q1C0Y0 CH60_YERPA
91.7 60 kDa chaperonin A7FN01 CH60_YERP3
92.3 60 kDa chaperonin Q6D9J0 CH60_PECAS
93.0 60 kDa chaperonin B2VL84 CH60_ERWT9
93.6 60 kDa chaperonin O66204 CH60_SERFI
94.2 60 kDa chaperonin O66206 CH60_SERMA
93.8 60 kDa chaperonin O66222 CH60_ERWAP
93.4 60 kDa chaperonin O66202 CH60_SERRU
93.6 60 kDa chaperonin C5BDK5 CH60_EDWI9
95.7 60 kDa chaperonin P95800 CH60_STEMA
94.4 60 kDa chaperonin O66218 CH60_PANAN
95.1 60 kDa chaperonin A8G8S7 CH60_SERP5
95.7 60 kDa chaperonin O66026 CH60_KLEPN
95.7 60 kDa chaperonin O66220 CH60_PECCC
96.0 60 kDa chaperonin O66210 CH60_KLEOX
96.0 60 kDa chaperonin C6DKC7 CH60_PECCP
96.0 60 kDa chaperonin O66200 CH60_ENTAG
96.6 60 kDa chaperonin A7MMC0 CH60_CROS8
96.0 60 kDa chaperonin O66214 CH60_RAOOR
96.2 60 kDa chaperonin A6TH53 CH60_KLEP7
96.2 60 kDa chaperonin B5Y368 CH60_KLEP3
96.4 60 kDa chaperonin O66190 CH60_ENTAS
96.2 60 kDa chaperonin O66198 CH60_KLEAK
96.8 60 kDa chaperonin A4W5N8 CH60_ENT38
97.4 60 kDa chaperonin O66194 CH60_PLUGE
97.0 60 kDa chaperonin O66196 CH60_LELAM
97.6 60 kDa chaperonin O66212 CH60_RAOPL
97.6 60 kDa chaperonin A8AMQ6 CH60_CITK8
97.7 60 kDa chaperonin O66192 CH60_KLUIN
98.7 60 kDa chaperonin B5R991 CH60_SALG2
98.7 60 kDa chaperonin A9MFR9 CH60_SALAR
98.9 60 kDa chaperonin P0A1D3 CH60_SALTY
98.9 60 kDa chaperonin P0A1D4 CH60_SALTI
98.9 60 kDa chaperonin B4TSC6 CH60_SALSV
98.9 60 kDa chaperonin B5BKF4 CH60_SALPK
98.9 60 kDa chaperonin C0Q6A2 CH60_SALPC
98.9 60 kDa chaperonin A9N3Z7 CH60_SALPB
98.9 60 kDa chaperonin Q5PL62 CH60_SALPA
98.9 60 kDa chaperonin B4TF80 CH60_SALHS
98.9 60 kDa chaperonin B5R005 CH60_SALEP
98.9 60 kDa chaperonin B5FRK2 CH60_SALDC
98.9 60 kDa chaperonin Q57GP7 CH60_SALCH
98.9 60 kDa chaperonin B5F2L0 CH60_SALA4
99.4 60 kDa chaperonin B7LLS5 CH60_ESCF3
99.6 60 kDa chaperonin Q0T9P8 CH60_ECOL5
100.0 60 kDa chaperonin Q3YUJ7 CH60_SHISS
100.0 60 kDa chaperonin P0A6F8 CH60_SHIFL
100.0 60 kDa chaperonin Q0SXD6 CH60_SHIF8
100.0 60 kDa chaperonin Q328C4 CH60_SHIDS
100.0 60 kDa chaperonin Q31T78 CH60_SHIBS
100.0 60 kDa chaperonin B2TY18 CH60_SHIB3
100.0 60 kDa chaperonin Q1R3B6 CH60_ECOUT
100.0 60 kDa chaperonin B1LQG4 CH60_ECOSM
100.0 60 kDa chaperonin B6I615 CH60_ECOSE
100.0 60 kDa chaperonin B7NG81 CH60_ECOLU
100.0 60 kDa chaperonin P0A6F5 CH60_ECOLI
100.0 60 kDa chaperonin B1ITQ5 CH60_ECOLC
100.0 60 kDa chaperonin P0A6F6 CH60_ECOL6
100.0 60 kDa chaperonin A8A7N9 CH60_ECOHS
100.0 60 kDa chaperonin B1XDP7 CH60_ECODH
100.0 60 kDa chaperonin C5A1D5 CH60_ECOBW
100.0 60 kDa chaperonin B7M8Q4 CH60_ECO8A
100.0 60 kDa chaperonin B7MSV9 CH60_ECO81
100.0 60 kDa chaperonin B7NTK2 CH60_ECO7I
100.0 60 kDa chaperonin B5Z2F2 CH60_ECO5E
100.0 60 kDa chaperonin P0A6F7 CH60_ECO57
100.0 60 kDa chaperonin B7LC02 CH60_ECO55
100.0 60 kDa chaperonin B7MKU8 CH60_ECO45
100.0 60 kDa chaperonin B7UPW3 CH60_ECO27
100.0 60 kDa chaperonin A7ZV12 CH60_ECO24
100.0 60 kDa chaperonin A1AJ51 CH601_ECOK1