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US 10,407,474 B2Granted
Protein surface remodeling
AssigneePresident and Fellows of Harvard College
InventorsDavid R. Liu, Kevin John Phillips, Michael S. Lawrence
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AssigneePresident and Fellows of Harvard College
InventorsDavid R. Liu, Kevin John Phillips, Michael S. Lawrence
Estimating the term…
Aggregation is a major cause of the misbehavior of proteins. A system for modifying a protein to create a more stable variant is provided. The method involves identifying non-conserved hydrophobic amino acid residues on the surface of a protein, suitable for mutating to more hydrophilic residues (e.g., charged amino acids). Any number of residues on the surface may be changed to create a variant that is more soluble, resistant to aggregation, has a greater ability to re-fold, and/or is more stable under a variety of conditions. The invention also provides GFP, streptavidin, and GST variants with an increased theoretical net charge created by the inventive technology. Kits are also provided for carrying out such modifications on any protein of interest.
A supercharged protein variant of a wild-type protein, wherein the supercharged protein variant comprises a modified primary amino acid sequence as compared to the wild-type sequence, resulting in a theoretical net charge on the supercharged protein variant of +10 to +52 at physiological pH, wherein the wild-type protein is selected from the group consisting of enzymes, transcription factors, DNA-binding proteins, and RNA-binding proteins.
Dependencies are inferred from the claim text.