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Expression, Solubilization, and Purification of Eukaryotic Borate Transporters
Published on: March 7, 2019
Characteristics affecting expression and solubilization of yeast membrane proteins
Michael A White1, Kathleen M Clark, Elizabeth J Grayhack
1Department of Biochemistry and Biophysics, University of Rochester Medical Center, Rochester, NY 14642, USA.
Journal of Molecular Biology
|November 3, 2006
Summary
Predicting membrane protein success is possible. Bioinformatic analysis of protein properties can identify candidates for overexpression and detergent solubilization, aiding structural studies.
Area of Science:
- Biochemistry
- Structural Biology
- Bioinformatics
Background:
- Membrane protein overexpression and solubilization are crucial for biochemical and structural analysis.
- Identifying predictive properties for successful membrane protein expression is essential.
Purpose of the Study:
- To identify properties of eukaryotic membrane proteins predictive of successful overexpression.
- To assess the detergent solubility of highly expressed yeast membrane proteins.
- To explore the utility of bioinformatic approaches for predicting membrane protein expression and solubility.
Main Methods:
- Analysis of expression levels for over 1000 predicted Saccharomyces cerevisiae membrane proteins.
- Statistical correlation analysis between expression levels and protein properties (size, hydrophobicity, transmembrane helices, amino acid composition).
- Detergent solubility testing of 122 highly expressed yeast membrane proteins using six common detergents.
Main Results:
- Statistically significant correlations found between high membrane protein expression and properties like size, hydrophobicity, and transmembrane helix count.
- High-level membrane protein expression positively correlated with the hydrophobicity of transmembrane segments, unlike soluble proteins.
- Most tested yeast membrane proteins were solubilized by a limited set of detergents, with solubility influenced by protein size, transmembrane helices, and segment hydrophobicity.
Conclusions:
- Bioinformatic approaches can predict membrane proteins amenable to overexpression and detergent solubilization.
- These methods can guide the selection of membrane proteins for structural and biochemical studies.
- Bioinformatics may aid in redesigning proteins with poor expression or solubility characteristics.
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