Peptide Linker for Transmembrane Protein Organelle Localization

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Solution Overview

Problem

Transmembrane proteins often face challenges in being smoothly localized to target organelles due to their high hydrophobicity, leading to aggregation or mislocalization to the endoplasmic reticulum.

Innovation Solution

The use of a peptide linker, either proline-rich or with an α-helix secondary structure, inserted between the transmembrane protein and an organelle localization signal, to facilitate the localization of transmembrane proteins to specific organelles by inhibiting intramolecular hydrophobic interactions.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If an organelle localization signal is imparted to a transmembrane protein, then the transmembrane protein should be localized to the target organelle, but the transmembrane protein aggregates between molecules or migrates to endoplasmic reticulum due to its high hydrophobicity

Engineering Contradiction:
Improvelocalization precisionVSAvoidlocalization efficiency
Core Design Contradiction:
Manufacturing precisionVSReliability

Solution Approach 1:

The patent introduces a peptide linker as an intermediary component between the transmembrane protein and the organelle localization signal. This linker acts as a mediator that prevents direct hydrophobic interactions between the transmembrane protein molecules, thereby preventing aggregation while still allowing the localization signal to function. The linker sequence (e.g., GGGGS repeats or proline-rich sequences) provides a hydrophilic spacer that maintains the spatial separation needed to prevent mislocalization to the endoplasmic reticulum.

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent segments the fusion protein into distinct functional modules: the transmembrane protein domain, the peptide linker domain, and the organelle localization signal domain. This segmentation allows each component to perform its specific function independently - the transmembrane protein maintains its membrane-spanning capability, the linker prevents aggregation, and the localization signal directs trafficking to the target organelle, thereby resolving the contradiction between maintaining protein function and achieving proper localization.

Inventive Principle:
Principle #1Segmentation

2Stability of the object's composition

If a transmembrane protein is made more hydrophobic to maintain membrane penetration capability, then membrane localization is improved, but aggregation between molecules increases and proper organelle localization decreases

Engineering Contradiction:
Improvemembrane localization stabilityVSAvoidorganelle localization precision
Core Design Contradiction:
Stability of the object's compositionVSManufacturing precision

Solution Approach 1:

The peptide linker serves as a hydrophilic intermediary that separates the hydrophobic transmembrane protein from the hydrophilic organelle localization signal. This mediator allows the transmembrane protein to maintain its hydrophobic character and membrane penetration capability without directly interacting with the localization signal, thereby preventing aggregation and ensuring precise organelle localization while preserving membrane stability.

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent applies local quality by creating distinct hydrophobic and hydrophilic regions within the fusion protein construct. The transmembrane protein region maintains its hydrophobic properties for membrane insertion, while the peptide linker region provides hydrophilic character to prevent aggregation, and the organelle localization signal region provides the necessary trafficking information. Each region has optimized local properties to fulfill its specific function without interfering with others.

Inventive Principle:
Principle #3Local quality

Data Source

PatentEP4549573A1Peptide linker and localization method for localizing transmembrane protein to target organelle, and localizing fusion protein
Publication Date: 2025.05.07 HAMAMATSU PHOTONICS KK
  • EP4549573A1 patent drawingFigure 1
  • EP4549573A1 patent drawingFigure 2
  • EP4549573A1 patent drawingFigure 3

AI summary

Disclosed are a highly rigid peptide linker inserted between a transmembrane protein and an organelle transport signal, a fusion protein containing the same, and a localization method that includes inserting a peptide linker.