Kinase Activity Measurement Using Effective Phosphorylation Site Counts
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Solution Overview
Problem
Conventional kinase activity measurements do not account for the variation in the number of phosphorylation sites of substrates, leading to inconsistencies in the production rate of phosphorylated amino acids and inaccurate activity assessments.
Innovation Solution
A method and apparatus that correlate the number of effective phosphorylation sites with substrate proteins, allowing for the estimation of kinase activity by quantifying the substrate protein amount and correcting the phosphorylation site count, using techniques like SPR and fluorescence labeling.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional kinase activity measurement methods are used, then the measurement process is simple, but the measurement precision is poor because the number of phosphorylation sites is not taken into consideration
Solution Approach 1:
The patent applies preliminary action by determining and storing the number of effective phosphorylation sites for each substrate protein before conducting kinase activity measurements. This pre-characterization data is stored in a database, allowing the measurement system to automatically retrieve and apply the correct phosphorylation site count during analysis, thereby improving precision without adding complexity to the actual measurement process
Solution Approach 2:
The patent introduces an intermediary computational layer that bridges the simple measurement process and the complex biochemical reality. A computer system acts as an intermediary, taking the measured phosphorylation levels and the pre-stored phosphorylation site counts to calculate normalized kinase activity values, thus resolving the contradiction between measurement simplicity and precision
2Measurement precision
If the number of phosphorylation sites is taken into consideration, then the kinase activity measurement precision is improved, but the device complexity increases due to additional data management requirements
Solution Approach 1:
The patent applies universality by creating a multi-functional system where the computer system serves multiple purposes: storing substrate protein information, managing phosphorylation site data, retrieving appropriate data during measurements, and performing normalized calculations. This universal data management approach consolidates multiple functions into a single integrated system, improving precision while managing complexity through consolidation rather than proliferation of separate components
Solution Approach 2:
The patent applies parameter changes by transforming the raw phosphorylation measurement data into normalized kinase activity values through mathematical processing. The system changes the parameter representation from absolute phosphorylation levels to normalized rates that account for the number of effective phosphorylation sites, thereby improving measurement precision while using standard computational methods that do not significantly increase system complexity
Data Source
AI summary
A method for measuring kinase activity taking into consideration the number of phosphorylation sites. The method for measuring kinase activity includes making the number of effective phosphorylation sites correspond to a substrate protein; acquiring the amount of the substrate protein on a base; and estimating the kinase activity based on the number of effective phosphorylation sites corresponding to the substrate protein on the base and the acquired amount of the substrate protein on the base.