Chimeric Olfactory Receptor cAMP Production
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Chimeric olfactory receptors unexpectedly decrease the production of cAMP, contrary to expectations, and there is a need for a method to increase cAMP production effectively.
Innovation Solution
A method involving a chimeric olfactory receptor with specific modifications, including an amino acid sequence derived from a mouse olfactory receptor and a beta-1 adrenergic receptor, where the IC4 domain is substituted and the N-terminal is modified with a Rho tag and myc epitope tag, is used in a reaction system with a lipid bilayer membrane and adenylate cyclase to enhance cAMP production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a chimeric olfactory receptor incorporating parts of beta-1 adrenergic receptor is used to increase cAMP production, then measurement accuracy should improve, but the chimeric olfactory receptor actually decreases cAMP production
Solution Approach 1:
The patent applies parameter changes by modifying the intracellular domain structure of the olfactory receptor. Specifically, it replaces the intracellular C-terminal domain (IC4) of the mouse olfactory receptor with the corresponding domain from beta-1 adrenergic receptor, and further optimizes by introducing specific point mutations (S371A, S375A, S378A) in the IC4 domain to restore cAMP production capability while maintaining the chimeric structure's other benefits
Solution Approach 2:
The patent applies local quality by making targeted modifications only to specific regions of the receptor protein. Instead of replacing the entire receptor or making global changes, it focuses modifications on the intracellular C-terminal domain (IC4) and specific serine residues within that domain, leaving the rest of the receptor structure intact to maintain its odorant binding and signaling capabilities
2Productivity
If the IC4 domain of olfactory receptor is substituted with beta-1 adrenergic receptor IC4 domain, then cAMP production should increase, but point mutations in IC3 and IC4 domains significantly decrease cAMP production
Solution Approach 1:
The patent applies parameter changes by introducing specific point mutations (S371A, S375A, S378A) in the IC4 domain of the chimeric receptor. These mutations change the phosphorylation sites from serine to alanine, preventing inhibitory phosphorylation and thereby restoring and stabilizing cAMP production capability in the chimeric receptor system
Solution Approach 2:
The patent applies copying by creating a modified version of the beta-1 adrenergic receptor IC4 domain that is copied into the olfactory receptor framework. The sequence from beta-1 adrenergic receptor is copied and then further optimized with specific mutations to achieve the desired functional outcome of increased stable cAMP production
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 modified chimeric olfactory receptor system significantly increases cAMP production, improving measurement accuracy and sensitivity to chemical substances.
Implementation Method 1
The olfactory receptor is a trimeric G protein-coupled receptor (hereinafter, referred to as 'GPCR'). The odor molecule binds to the olfactory receptor. The binding separates the trimeric G protein into the alpha subunit (Gαolf) and a beta-gamma complex. The separated Gαolf activates the adenylate cyclase (AC). The activated adenylate cyclase (AC) converts adenosine triphosphate (ATP) into cyclic adenosine monophosphate (cAMP).
Implementation Method 2
The cell membrane is mainly composed of a lipid bilayer membrane. The lipid bilayer membrane has a structure of two layers each consisting of phospholipid molecules arranged in a high density.
Implementation Method 3
The cyclic adenosine monophosphate (cAMP) activates an ion channel, more particularly, for example, a cyclic nucleotide gated ion channel (CNG). The activation allows an ion to be transported from the inside of the cell to the outside of the cell, or from the outside of the cell to the inside of the cell. The degree of the transport of the ion can be measured as an electric signal.
Data Source
AI summary
A method for producing cAMP using a chimeric olfactory receptor. The method includes a step of preparing a reaction system comprising a first layer, a lipid bilayer membrane, and a second layer, and a step of supplying a chemical substance which stimulates the chimeric olfactory receptor to the first layer so as to produce the cAMP from ATP. The lipid bilayer membrane includes the chimeric olfactory receptor and adenylate cyclase. The chimeric olfactory receptor penetrates the lipid bilayer membrane. The second layer contains ATP and a G protein. The G protein is placed in the vicinity of one end of the chimeric olfactory receptor. The chimeric olfactory receptor is derived from a mouse olfactory receptor and the N-terminal of the chimeric olfactory receptor is modified with an amino acid sequence.