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Updated: Aug 2, 2025

Specificity Analysis of Protein Lysine Methyltransferases Using SPOT Peptide Arrays

Specificity Analysis of Protein Lysine Methyltransferases Using SPOT Peptide Arrays

Published on: November 29, 2014

Identifying Key Residues in Lysine Decarboxylase for Soluble Expression Using Consensus Design Soluble Mutant

Jin Young Kim¹, Gyeong-Guk Park¹, Eun-Jung Kim^2,3

¹Interdisciplinary Program for Biochemical Engineering and Biotechnology, Seoul National University, Seoul 08826, Republic of Korea.

ACS Synthetic Biology

|April 18, 2023

Summary

This summary is machine-generated.

This study introduces ConsenSing, a hybrid method combining sequence analysis and experimental screening to improve protein solubility. It successfully identified mutations that significantly enhance the soluble expression of target proteins like LdcC.

Keywords:

consensus sequence fluorescence-activated cell sorting (FACS)lysine decarboxylase soluble expression split green fluorescence protein (GFP)

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Last Updated: Aug 2, 2025

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A Facile Protocol to Generate Site-Specifically Acetylated Proteins in Escherichia Coli

A Facile Protocol to Generate Site-Specifically Acetylated Proteins in Escherichia Coli

Published on: December 9, 2017

Area of Science:

Biochemistry
Protein Engineering
Computational Biology

Background:

Deep learning models predict protein solubility but experimental validation is challenging.
Rapidly confirming computational predictions is crucial for protein engineering success.

Purpose of the Study:

To develop a hybrid computational and experimental approach for predicting and validating protein solubility-improving mutations.
To identify key residues that enhance soluble protein expression using a novel screening strategy.

Main Methods:

Developed ConsenSing (Consensus design Soluble Mutant Screening), a hybrid approach using sequence-based analysis for hot spot prediction.
Constructed a compact mutant library using Darwin assembly for efficient empirical screening.
Utilized split GFP as a reporter system for validating soluble expression.

Main Results:

Identified multiple mutants of *Escherichia coli* lysine decarboxylase (LdcC) with significantly increased soluble expression.
Pinpointed a single critical residue responsible for enhanced LdcC soluble expression.
Unveiled the mechanism by which a single mutation improves protein solubility and expression.

Conclusions:

ConsenSing effectively links computational predictions with experimental validation for protein solubility enhancement.
Following natural evolutionary paths can guide single-residue mutations to significantly improve protein solubility and expression.
The study provides a validated framework for rapid protein engineering to boost soluble protein production.