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Updated: Jun 19, 2026
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Evaluation of Protein–Protein Interactions using an On-Membrane Digestion Technique
Published on: July 19, 2019
Quantification of membrane proteins using nonspecific protease digestions
Maria Bendz1, Mirja Carlsson Möller, Giorgio Arrigoni
1Protein Technology, Department of Immunotechnology, CREATE Health, Lund University, Sweden.
Journal of Proteome Research
|October 23, 2009
Summary
We developed a new mass spectrometry method to identify and quantify membrane proteins. This technique reveals changes in Bacillus subtilis membrane protein expression when grown with or without glucose.
Area of Science:
- Proteomics
- Analytical Chemistry
- Microbiology
Background:
- Membrane proteins play crucial roles in cellular functions but are challenging to study.
- Existing methods for membrane protein analysis often face limitations in scope and sensitivity.
Purpose of the Study:
- To develop and validate a novel mass spectrometry-based method for comprehensive membrane protein identification and quantification.
- To apply this method to investigate differential membrane protein expression in Bacillus subtilis under varying nutritional conditions.
Main Methods:
- Proteins were isolated using a modified SDS-PAGE protocol.
- High-pH digestion was performed using the low-specificity protease Proteinase K.
- Peptides were labeled with a fragmentation-directing isotope tag for mass spectrometry analysis.
Main Results:
- The method successfully identified and quantified membrane proteins from Bacillus subtilis.
- Significant differences in membrane protein expression were observed between cells grown in the presence and absence of glucose.
Conclusions:
- The developed mass spectrometry approach offers a robust platform for membrane proteome analysis.
- This method provides insights into metabolic regulation of membrane protein expression in bacteria.
