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Proteomics approach to identifying ATP-covalently modified proteins
Paul G Besant1, Michael V Lasker, Cuong D Bui
1Department of Medicine, University of California-San Francisco, Parnassus, U-426, San Francisco, California 94143-0724, USA. pbesant@cyllene.uwa.edu.au
Journal of Proteome Research
|March 5, 2004
Summary
Researchers identified proteins that bind ATP, suggesting functional similarity. This proteomics method screens for proteins with ATP-binding capabilities for further analysis.
Area of Science:
- Biochemistry
- Proteomics
- Molecular Biology
Background:
- Proteins with similar functions often share conserved properties.
- Adenosine triphosphate (ATP) binding is a critical function for many proteins.
- Identifying proteins based on ATP binding can reveal functional relationships.
Purpose of the Study:
- To investigate functionally similar proteins by assessing their ability to remain bound to ATP.
- To develop a screening method for identifying proteins with ATP-binding capabilities.
Main Methods:
- Utilized two-dimensional gel electrophoresis (2D-PAGE) combined with capillary liquid chromatography-on-line mass spectrometry (LC-MS).
- Screened mammalian and E. coli protein extracts for ATP-bound proteins.
- Validated findings by testing commercially purified proteins for ATP binding using radiolabeled ATP (α-32P ATP).
Main Results:
- Identified several mammalian and E. coli proteins that appear to covalently bind ATP.
- Confirmed ATP-binding capacity in purified proteins initially identified through the proteomic screen.
- Demonstrated the feasibility of using ATP binding as a criterion for protein similarity screening.
Conclusions:
- The described proteomics approach serves as an effective initial screening method.
- This technique can identify functionally related proteins for subsequent detailed investigation.
- ATP binding is a valuable characteristic for classifying and identifying proteins with similar functions.