Ankyrin Repeat Domain Thermostability via Position 23 Mutation
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
There is a need to further improve the thermostability of ankyrin repeat domains, which are already stable but can benefit from additional enhancements for better performance in various biological applications.
Innovation Solution
The ankyrin repeat domain is engineered by mutating position 23 of the internal ankyrin repeat adjacent to the N-terminal capping module to an amino acid residue of the leucine class, such as leucine or isoleucine, to enhance the interaction between the N-terminal capping module and the internal ankyrin repeat, thereby increasing the overall stability of the domain.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If consensus ankyrin repeat motifs are used to design recombinant ankyrin repeat scaffolds, then the structural integrity and basic stability are achieved, but the thermostability can be further improved
Solution Approach 1:
The patent applies local quality by introducing specific amino acid substitutions at particular positions within the ankyrin repeat domain rather than changing the entire sequence. Positions 15, 17, 22, and 23 are targeted for mutation to hydrophobic or leucine-class residues, creating localized improvements in thermostability while preserving the overall consensus structure and function.
Solution Approach 2:
The patent changes physical-chemical parameters of the protein sequence by substituting specific residues with those having different properties (hydrophobicity, size, charge). These parameter changes at key positions enhance the hydrophobic core packing and inter-repeat interactions, thereby improving thermostability without fundamentally altering the protein's identity or function.
2Ease of manufacture
If surface-exposed polar residues are added to terminal consensus ankyrin repeats, then solubility issues are mitigated, but the thermostability may be compromised
Solution Approach 1:
The patent maintains the solubility-enhancing polar residues at terminal positions while applying localized mutations at internal positions (15, 17, 22, 23) that specifically target thermostability improvement. This spatial separation allows both solubility and thermostability to be optimized independently at different locations within the same protein structure.
Data Source
AI summary
Described herein are proteins comprising an ankyrin repeat domain with a mutation in the internal ankyrin repeat that is adjacent to the N-terminal capping module, in particular a mutation at position 23 of said internal ankyrin repeat, as well as related products and methods.