The Allosteric Transition of GroEL Induced by Metal Fluoride–ADP Complexes

Inobe, Tomonao; Kikushima, Kenji; Arai, Munehito; Kuwajima, Kunihiro

doi:10.1016/s0022-2836(03)00409-1

Cited by 20 publications

(28 citation statements)

References 36 publications

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“…In the present study, we found that ADP ϩ BeF x , which is thought to mimic the ATP-bound state (19,20,24), induced the conformation on ␣-chaperonin with one ring closed. In contrast, it was shown that ADP ϩ AlF x , an analog of the trigonal-bipyramidal transition state in ATP hydrolysis, induced the lid closure of CCT (13).…”

Section: Discussionsupporting

confidence: 52%

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Characterization of Archaeal Group II Chaperonin-ADP-Metal Fluoride Complexes

Iizuka

Yoshida

Ishii

et al. 2005

Journal of Biological Chemistry

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Group II chaperonins, found in Archaea and in the eukaryotic cytosol, act independently of a cofactor corresponding to GroES of group I chaperonins. Instead, the helical protrusion at the tip of the apical domain forms a built-in lid of the central cavity. Although many studies on the lid's conformation have been carried out, the conformation in each step of the ATPase cycle remains obscure. To clarify this issue, we examined the effects of ADP-aluminum fluoride (AlF x ) and ADP-beryllium fluoride (BeF x ) complexes on ␣-chaperonin from the hyperthermophilic archaeum, Thermococcus sp. strain KS-1. Biochemical assays, electron microscopic observations, and small angle x-ray scattering measurements demonstrate that ␣-chaperonin incubated with ADP and BeF x exists in an asymmetric conformation; one ring is open, and the other is closed. The result indicates that ␣-chaperonin also shares the inherent functional asymmetry of bacterial and eukaryotic cytosolic chaperonins. Most interestingly, addition of ADP and BeF x induced ␣-chaperonin to encapsulate unfolded proteins in the closed ring but did not trigger their folding. Moreover, ␣-chaperonin incubated with ATP and AlF x or BeF x adopted a symmetric closed conformation, and its functional turnover was inhibited. These forms are supposed to be intermediates during the reaction cycle of group II chaperonins.Chaperonins, one of the most well studied molecular chaperones, are ubiquitous and indispensable proteins that are involved in protein folding in the cell. They assist in protein folding reactions both in vivo and in vitro by binding unfolded proteins within the central cavity of the double ring structure (1, 2). Based on protein sequence and structural features, chaperonins fall into two groups (2, 3). Group I chaperonins are found in bacteria, endosymbiotic organelles (mitochondria and chloroplasts), and only subsets of Archaea (4, 5). The most extensively characterized member is the Escherichia coli chaperonin GroEL and its partner GroES. GroEL-mediated protein folding requires GroES as a lid of the GroEL cavity. GroEL is functionally asymmetrical and is referred to as a "two-stroke engine," with one ring binding ATP and GroES, and simultaneously acting as the place where protein folding occurs, followed by an identical reaction on the opposite ring (6 -8). In contrast, group II chaperonins, found in Archaea (called thermosome) and the eukaryotic cytosol (known as CCT or TCP-1 ring complex), work without the cofactor corresponding to GroES. The helical protrusion at the tip of the apical domain substitutes for the cofactor as a built-in lid of the central cavity. Many studies on the change of the lid's conformation coupled to the binding and hydrolysis of nucleotides have been carried out. It was found that ATP drives the conformational change of group II chaperonins from the open lid, substrate binding conformation to the closed lid conformation to encapsulate unfolded protein in the central cavity (9 -15). However, the details of the ATP-driven reaction in g...

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Section: Discussionsupporting

confidence: 52%

“…19 -21). They have also been used to study the functional cycle of chaperonins (9,13,(22)(23)(24)(25)(26). The crystal structure of the Thermoplasma chaperonin complexed with ADP and AlF x (Protein Data Bank code 1A6E) provided the enzymatic information on the mechanism of ATP hydrolysis (9).…”

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confidence: 99%

Characterization of Archaeal Group II Chaperonin-ADP-Metal Fluoride Complexes

Iizuka

Yoshida

Ishii

et al. 2005

Journal of Biological Chemistry

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“…AlF x was also tested but, unlike BeF x , several minutes were required to gain the maximum inhibition (data not shown). Another phosphate analogue, NaVO 4 , did not show an inhibitory effect on GroEL ATPase, as reported previously (18,19). Therefore, we focused on the effect of BeF x .…”

Section: Inhibition Of Groel Atpase Turnover By Bef X -mentioning

confidence: 63%

“…Indeed, recent x-ray crystallography of the 1:1 GroEL-GroES formed in AlF x showed that AlF 3 is coordinated to ADP at each of the catalytic sites in the cis-ring of the complex (19). X-ray crystallography, as well as x-ray scattering analysis, also suggested that the 1:1 GroEL-GroES complexes formed in metal fluorides resemble the usual 1:1 GroEL-GroES complex formed in the absence of metal fluorides (18,19). We recently reported that ADP with minimum ATP contamination can mediate the binding of GroES to the substrate protein-free GroEL but not to the substrate proteinloaded GroEL (21).…”

Section: Discussionmentioning

confidence: 95%

“…To investigate what happens if P i release is prevented, we have adopted fluoroberyllate (BeF x ) 1 that can mimic the hydrolysis-product P i or ␥-phosphate part of the enzyme-bound nucleotide and can form a stable complex with ADP (or GDP) in a variety of ATP (or GTP)-utilizing enzymes (16) including the chaperonin family (17)(18)(19). When we added BeF x , the GroEL functional cycle was stopped immediately and a symmetric 1:2 GroEL-GroES complex 2 was formed in which both two GroEL rings contained a folding competent encapsulated substrate protein.…”

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BeF Stops the Chaperonin Cycle of GroEL-GroES and Generates a Complex with Double Folding Chambers

Taguchi

Tsukuda

Motojima

et al. 2004

Journal of Biological Chemistry

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Coupling with ATP hydrolysis and cooperating with GroES, the double ring chaperonin GroEL assists the folding of other proteins. Here we report novel GroEL-GroES complexes formed in fluoroberyllate (BeFx) that can mimic the phosphate part of the enzyme-bound nucleotides. In ATP, BeFxstops the functional turnover of GroEL by preventing GroES release and produces a symmetric 1:2 GroEL-GroES complex in which both GroEL rings contain ADP·BeFxand an encapsulated substrate protein. In ADP, the substrate protein-loaded GroEL cannot bind GroES. In ADPplusBeFx, however, it can bind GroES to form a stable 1:1 GroEL-GroES complex in which one of GroEL rings contains ADP·BeFxand an encapsulated substrate protein. This 1:1 GroEL-GroES complex is converted into the symmetric 1:2 GroEL-GroES complex when GroES is supplied in ATPplusBeFx. Thus, BeFxstabilizes two GroEL-GroES complexes; one with a single folding chamber and the other with double folding chambers. These results shed light on the intermediate ADP·Pinucleotide states in the functional cycle of GroEL.

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The allosteric transition of the chaperonin groel fromescherichia coli as studied by solution X-ray scattering

2006

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This is a short review article of our recent studies on the ATP-induced, allosteric conformational transition of the chaperonin GroEL complex by solution X-ray scattering. We used synchrotron X-ray scattering with a twodimensional, charge-coupled, device-based X-ray detector to study (1) the specificity of the chaperonin GroEL for its ligand that induced the allosteric transition, and (2) the identification of the allosteric transition of GroEL in its complicated kinetics induced by ATP. Due to the dramatically increased sensitivity of the X-ray scattering technique based on the use of the two dimensional X-ray detector and synchrotron radiation, different allosteric conformational states of GroEL populated under different conditions were clearly distinguished from each other. It was concluded that solution X-ray scattering is an extremely powerful tool for investigating the equilibrium and kinetics of cooperative conformational transitions of oligomeric protein complex, especially when combined with other spectroscopic techniques such as fluorescence spectroscopy.

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The Allosteric Transition of GroEL Induced by Metal Fluoride–ADP Complexes

Cited by 20 publications

References 36 publications

Characterization of Archaeal Group II Chaperonin-ADP-Metal Fluoride Complexes

Characterization of Archaeal Group II Chaperonin-ADP-Metal Fluoride Complexes

BeF Stops the Chaperonin Cycle of GroEL-GroES and Generates a Complex with Double Folding Chambers

The allosteric transition of the chaperonin groel fromescherichia coli as studied by solution X-ray scattering

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