BBRC manuscript

Original Text: BBRC manuscript


Molecular chaperones appear to have evolved to facilitate protein folding in the cell through the entrapment of folding intermediates on the interior of a large cavity formed between GroEL and its co-chaperonin GroES by binding newly synthesized or non-native polypeptides through hydrophobic interactions and prevent their aggregation. Some proteins do not interact with GroEL, hence even though they are aggregation prone, cannot be assisted by GroEL for their folding. Here, we have attempted engineering of these non-substrate proteins to convert them to the substrate for GroEL, without compromising their function. We have used computational biology approaches to generate mutants of the selected proteins by selectively mutating residues in the hydrophobic patch, similar to GroES mobile loop region which is responsible for interaction with GroEL, and compared with the wild counterparts and calculated their instability and aggregation propensities. The energies of the newly designed mutants were computed through Molecular Dynamics simulations. We observed increased aggregation propensity of some of the mutants formed after replacing charged amino acid residues with hydrophobic ones in the well defined hydrophobic patch, raising possibility of their binding ability to GroEL. The newly generated mutants may provide potential substrates for Chaperonin GroEL, which can be experimentally generated and tested for their tendency of aggregation, interactions with GroEL and possibility of chaperone assisted folding to produce functional protein.

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Revised Text:

Molecular chaperones appear to have evolved to facilitate protein folding in the cell through the entrapment of folding intermediates on the interior of a large cavity formed between GroEL and its co-chaperonin GroES by binding newly synthesized or non-native polypeptides through hydrophobic interactions and preventing their aggregation.

Some proteins do not interact with GroEL, so even though they are aggregation prone, they cannot be assisted by GroEL for their folding. Here, we have attempted engineering of these non-substrate proteins to convert them to the substrate for GroEL, without compromising their functions. We have used computational biology approaches to generate mutants of the selected proteins by selectively mutating residues in the hydrophobic patch. This is similar to the GroES mobile loop region which is responsible for interaction with GroEL. These were compared to their wild counterparts and their instability and aggregation propensities were calculated.

The energies of the newly designed mutants were computed through Molecular Dynamics simulations. We observed increased aggregation propensity of some of the mutants formed after replacing charged amino acid residues with hydrophobic ones in the well defined hydrophobic patch. This raised the possibility of their binding ability to GroEL.

The newly generated mutants may provide potential substrates for Chaperonin GroEL, which can be experimentally generated and tested for their tendency of aggregation, interactions with GroEL and possibility of chaperone assisted folding to produce functional proteins.


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