Amino Acid Binding Protein Sequencing via Segmentation
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Solution Overview
Problem
Current methods for determining large-scale protein sequencing information from biological samples face challenges due to the complexity of protein structure, composition, and modification.
Innovation Solution
The application provides methods and compositions for determining amino acid sequence information from polypeptides using amino acid binding proteins, which can be labeled and used in sequencing reactions to identify the relative position of amino acids and sequence polypeptides.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional protein sequencing methods are used, then large-scale protein sequencing information can be obtained, but the complexity of protein structure, composition, and modification makes determination difficult and imprecise
Solution Approach 1:
The polypeptide is divided into individual amino acid units through sequential cleavage at the N-terminus. Each amino acid is released and identified separately, converting the complex problem of determining entire protein sequences into manageable single-amino-acid identification steps, thereby improving measurement precision despite structural complexity
Solution Approach 2:
Aminoxyl radicals serve as intermediary species that facilitate selective labeling of terminal amino acids. These radicals mediate the interaction between the polypeptide and detection reagents, enabling precise identification of amino acid sequences without being affected by the overall complexity of protein structure and modifications
2Manufacturing precision
If labeled amino acid binding proteins are used for sequencing, then single polypeptide molecule sequencing is enabled, but the complexity of labeling and reaction composition increases
Solution Approach 1:
The amino acid binding protein is designed with multiple functions: it selectively binds to specific amino acid residues, carries a detectable label for identification, and can be used in sequential reaction cycles. This multi-functionality reduces the need for multiple different reagents, simplifying the overall reaction composition while maintaining high sequencing precision
Solution Approach 2:
The binding affinity and label characteristics of the amino acid binding protein are optimized to operate under specific reaction conditions. By controlling parameters such as pH, temperature, and label fluorescence properties, the system achieves precise sequencing while managing the complexity of the reaction composition through parameter optimization rather than additional components
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
This approach allows for the determination of amino acid sequence information from single polypeptide molecules, enabling precise sequencing and identification of proteins in complex samples.
Implementation Method 1
the one or more labels comprise a luminescent label
Implementation Method 2
the biotinylation sequence comprises two biotin ligase recognition sequences oriented in tandem
Data Source
AI summary
Aspects of the application provide methods of identifying and sequencing proteins, polypeptides, and amino acids, and compositions useful for the same. In some aspects, the application provides amino acid recognition molecules, such as amino acid binding proteins and fusion polypeptides thereof. In some aspects, the application provides amino acid recognition molecules comprising a shielding element that enhances photostability in polypeptide sequencing reactions.