Nucleic Acid Tether for Enzyme Co-localization
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Solution Overview
Problem
Current methods for co-localizing enzymes with their substrates are inefficient, often requiring enzyme modification and resulting in reduced activity, and lack simple and reliable ways to tether enzymes to their targets, limiting reaction rates and product yields.
Innovation Solution
The use of tethering mechanisms, specifically linking enzymes to substrates or surfaces using nucleic acid hybridization, which allows for efficient co-localization of enzymes with their substrates, maintaining enzymatic activity, and reducing protein consumption and loss during reactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If enzymes are immobilized on a support to co-localize with substrate, then enzyme co-localization with substrate is improved, but enzyme activity is reduced
Solution Approach 1:
A nucleic acid tether acts as an intermediary component that links the enzyme to the substrate or surface. The tether comprises a first end that links to the enzyme and a second end that hybridizes to a nucleic acid on the substrate or surface, enabling co-localization without direct immobilization of the enzyme on a support structure.
2Ease of manufacture
If enzymes are modified for immobilization, then enzyme tethering capability is improved, but technical complexity and time consumption increase
Solution Approach 1:
The nucleic acid tether serves as a mediator that eliminates the need for complex enzyme modification. The tether can be synthesized independently and then hybridized to both the enzyme (via affinity tags or chemical conjugation) and the substrate nucleic acid, simplifying the overall process.
Solution Approach 2:
The nucleic acid tether is prepared in advance with specific sequences designed for hybridization to substrate nucleic acids. This preliminary preparation of the tether component separates the complexity of nucleic acid synthesis from enzyme handling, allowing enzymes to be used in their native or minimally modified state.
3Stability of the object's composition
If conventional immobilization methods are used, then enzyme co-localization is achieved, but reaction efficiency and product yield decrease
Solution Approach 1:
The nucleic acid tether provides a flexible linkage that allows the enzyme to remain positioned near the substrate while maintaining freedom of movement for catalytic activity. This intermediary approach preserves the enzyme's natural function while achieving co-localization, thereby maintaining high reaction rates and product yields.
Solution Approach 2:
The nucleic acid tether provides dynamic flexibility rather than rigid immobilization. The tether can flex and move, allowing the enzyme to adopt optimal orientations for catalysis while remaining co-localized with the substrate, thus maintaining high productivity.
Data Source
AI summary
In some embodiments, the disclosure relates generally to methods as well as related compositions, systems, kits and apparatus comprising linking proteins to target compounds and/or to locations of interest using tethers. For example, the tether can be used to link the protein to a target compound, for example, to link an enzyme to a substrate. Similarly, the tether can be used to link the protein at or near a desired location on a surface. In one group of embodiments, the tether includes a polynucleotide and the target compound or location on the surface includes another polynucleotide that is capable of hybridizing to the tether. In such embodiments, the tether can be used to link the protein to the target compound or location using nucleic acid hybridization.