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• W2047650175 abstract "Noncovalently stacked tetramethylrhodamine (TMR) dimers have been used to both report and perturb the allosteric equilibrium in GroEL. A GroEL mutant (K242C) has been labeled with TMR, close to the peptide-binding site in the apical domain, such that TMR molecules on adjacent subunits are able to form dimers in the T allosteric state. Addition of ATP induces the transition to the R state and the separation of the peptide-binding sites, with concomitant unstacking of the TMR dimers. A statistical analysis of the spectra allowed us to compute the number and orientation of TMR dimers per ring as a function of the average number of TMR molecules per ring. The TMR dimers thus serve as quantitative reporter of the allosteric state of the system. The TMR dimers also serve as a surrogate for substrate protein, substituting in a more homogeneous, quantifiable manner for the heterogeneous intersubunit, intraring, noncovalent cross-links provided by the substrate protein. The characteristic stimulation of the ATPase activity by substrate protein is also mimicked by the TMR dimers. Using an expanded version of the nested cooperativity model, we determine values for the free energy of the TT to TR and TR to RR allosteric equilibria to be 27 ± 11 and 46 ± 2 kJ/mol, respectively. The free energy of unstacking of the TMR dimers was estimated at 2.6 ± 1.0 kJ/mol dimer. These results demonstrate that GroEL can perform work during the T to R transition, supporting the iterative annealing model of chaperonin function." @default.
• W2047650175 created "2016-06-24" @default.
• W2047650175 creator A5015387142 @default.
• W2047650175 creator A5054928202 @default.
• W2047650175 date "2013-05-30" @default.
• W2047650175 modified "2023-10-01" @default.
• W2047650175 title "Measuring how much work the chaperone GroEL can do" @default.
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• W2047650175 doi "https://doi.org/10.1073/pnas.1307837110" @default.
• W2047650175 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/3703975" @default.
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• W2047650175 hasPublicationYear "2013" @default.
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