ClpS2 Mutant Binders for Selective Amino Acid Detection
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Solution Overview
Problem
Current technologies face challenges in achieving high-fidelity, sequential recognition and detection of specific amino acids in peptide sequences, particularly due to lack of selectivity and affinity in existing N-End Rule Pathway adaptor proteins like ClpS.
Innovation Solution
Development of an amino acid-specific binder with novel sequence variants of Agrobacterium tumefaciens ClpS2, which exhibits enhanced affinity and selectivity for amino acids like phenylalanine, tryptophan, and tyrosine, enabling selective binding and identification of these amino acids.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If existing N-End Rule Pathway adaptor proteins like ClpS are used, then the system can recognize N-terminal amino acids, but the selectivity and affinity for specific amino acids are insufficient
Solution Approach 1:
The patent applies parameter changes by systematically mutating specific amino acid residues in the ClpS2 protein sequence to optimize binding properties. Variants such as A34P, R35R, and E36E are engineered to enhance both selectivity for specific N-terminal amino acids and binding affinity, directly resolving the contradiction between measurement precision and reliability
Solution Approach 2:
The invention implements local quality by focusing mutations on specific residues (positions 34-36) within the binding pocket of ClpS2. This localized modification approach allows the protein to maintain overall structure while achieving enhanced selectivity and affinity at the critical binding interface, addressing the technical contradiction without compromising overall protein function
2Measurement precision
If existing ClpS proteins are used, then amino acid recognition is possible, but high-fidelity sequential recognition and detection cannot be achieved
Solution Approach 1:
The patent employs copying by creating multiple variant versions of the ClpS2 protein, each optimized for detecting specific amino acids. These copied and modified protein variants can be used in parallel or sequentially to achieve high-fidelity detection of multiple amino acids, maintaining detection fidelity while managing system complexity through modular design
Solution Approach 2:
The invention applies segmentation by dividing the amino acid detection task into multiple steps, where different ClpS2 variants with specific mutational profiles are used to detect different amino acids sequentially. This segmented approach enables high-fidelity detection of complex peptide sequences while keeping each individual detection step relatively simple
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The amino acid-specific binder achieves high-fidelity, sequential recognition and detection of specific amino acids, overcoming the limitations of existing technologies by providing enhanced selectivity and affinity, thereby determining the sequence or fingerprint of amino acids in peptides or proteins.
Implementation Method 1
an amino acid-specific binder for selectively binding to an amino acid in an analyte
Data Source
AI summary
An amino acid-specific binder selectively binds to a binding amino acid. A binder complex selectively identifies the binding amino acid and includes an adjunct attached to the amino acid-specific binder. The adjunct includes a taggant, protein, substrate, or chemical modifier. Selectively identifying an N-terminal amino acid includes anchoring a C-terminal end; contacting an N-terminal amino acid of the anchored analyte with the binder complex; selectively binding when the N-terminal amino acid includes the binding amino acid; producing, by the taggant of the tagged complex, a taggant signal; detecting the taggant signal; and identifying the N-terminal amino acid based on the taggant signal.