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• W2316494629 abstract "The fundamental principles that govern monomer folding are believed to be congruent with those of protein oligomers. However, the effects of protein assembly during the folding reaction can result in a series of complex transitions that are considerably more challenging to deconvolute. Here we developed the experimental protein folding mechanism for the lactose repressor (LacI), for both the dimeric and the tetrameric states, using equilibrium unfolding and kinetic experiments, and by leveraging the previously reported monomer folding landscape. Reaction details for LacI oligomers were observed by way of circular dichroism, intrinsic fluorescence, and Förster resonance energy transfer (FRET) and as a function of protein concentration. In general, the dimer and tetramer are four-phase folding reactions in which the first three transitions are tantamount to the folding of constituent monomers. The final reaction phase of the LacI dimer can be attributed to protein assembly, based on the concentration dependence of the observed folding rates and intermolecular FRET measurements. Unlike the dimer, the latter reaction phase of the LacI tetramer is not dependent on protein concentration, likely because of a strong tethering of the monomers, which simplifies the folding reaction by eliminating an explicit protein assembly phase. Finally, folding of the LacI dimer and tetramer was assessed in the presence of polyethylene glycol to rule out inert molecular crowding as the driving force for the protein folding reaction; in addition, these data provide insight into the folding mechanism in vivo." @default.
• W2316494629 created "2016-06-24" @default.
• W2316494629 creator A5051928700 @default.
• W2316494629 creator A5058038414 @default.
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• W2316494629 date "2012-09-10" @default.
• W2316494629 modified "2023-09-25" @default.
• W2316494629 title "Lactose Repressor Experimental Folding Landscape: Fundamental Functional Unit and Tetramer Folding Mechanisms" @default.
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• W2316494629 doi "https://doi.org/10.1021/bi300545f" @default.
• W2316494629 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/22931511" @default.
• W2316494629 hasPublicationYear "2012" @default.
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