Mutational probing of protein aggregates to design aggregation‐resistant proteins

Mutational probing of protein aggregates to design aggregation‐resistant proteins

Characterization of amorphous protein aggregates may offer insights into the process of aggregation. Eleven single amino acid mutants of lipase (LipA of Bacillus subtilis) were subjected to temperature‐induced aggregation, and the resultant aggregates were characterized for recovery of activity in t...

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Bibliographic Details
Published in:FEBS open bio Vol. 6; no. 2; pp. 126 - 134
Main Authors: Kamal, Mohamad Zahid, Kumar, Virender, Satyamurthi, Kundarapu, Das, Kushal Kumar, Rao, Nalam Madhusudhana
Format: Journal Article
Language:English
Published: England John Wiley & Sons, Inc 01-02-2016
John Wiley and Sons Inc
Wiley
Subjects:
Aggregates
aggregation resistance
Amino acids
Enzymes
Heat
Insulin
intermediates
Kinetics
LipA protein
local unfolding
Mutation
mutations
Protein interaction
protein stability
Proteins
aggregation resistance
mutations
intermediates
local unfolding
protein stability
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Summary:Characterization of amorphous protein aggregates may offer insights into the process of aggregation. Eleven single amino acid mutants of lipase (LipA of Bacillus subtilis) were subjected to temperature‐induced aggregation, and the resultant aggregates were characterized for recovery of activity in the presence of guanidinium chloride (GdmCl). Based on activity recovery profiles of the aggregates, the mutants could be broadly assigned into four groups. By including at least one mutation from each group, a mutant was generated that showed an increase of ~ 10 °C in melting temperature (Tm) compared to the wild‐type and did not aggregate even at 75 °C. This method explores characterization of amorphous protein aggregates in the presence of GdmCl and helps in identifying mutations involved in protein aggregation.
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ISSN:2211-5463
2211-5463
DOI:10.1002/2211-5463.12003