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Updated: May 9, 2026
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Phosphopeptide Enrichment Coupled with Label-free Quantitative Mass Spectrometry to Investigate the Phosphoproteome in Prostate Cancer
Published on: August 2, 2018
Purification of prostatic acid phosphatase (PAP) for structural and functional studies
Annakaisa M Herrala1, Ileana B Quintero, Pirkko T Vihko
1Department of Clinical Chemistry, Institute of Clinical Medicine, University of Helsinki, Helsinki, Finland.
Methods in Molecular Biology (Clifton, N.J.)
|July 18, 2013
Summary
This study presents a robust purification method for prostatic acid phosphatase (PAP) using chromatography techniques. The purified PAP enabled the determination of its first reported three-dimensional structure.
Area of Science:
- Biochemistry
- Structural Biology
- Protein Purification
Background:
- High-scale protein purification is essential for various biochemical and structural studies.
- Prostatic acid phosphatase (PAP) is a key enzyme with implications in prostate cancer research.
- Previous methods may have limitations for obtaining sufficient quantities of highly pure PAP.
Purpose of the Study:
- To develop and describe an efficient, high-scale purification method for prostatic acid phosphatase (PAP).
- To obtain purified PAP suitable for structural determination.
- To facilitate further functional and structural investigations of PAP.
Main Methods:
- Purification of PAP utilizing a combination of anion exchange chromatography, L-(+)-tartrate affinity chromatography, and gel filtration chromatography.
- Monitoring protein concentration and acid phosphatase activity throughout the purification process.
- Utilizing the p-nitrophenyl phosphate assay to identify fractions with optimal enzyme activity.
Main Results:
- A successful purification protocol for prostatic acid phosphatase (PAP) was established.
- The method yielded sufficient quantities of purified PAP for downstream applications.
- The purified protein was utilized to determine the first reported three-dimensional structure of PAP.
Conclusions:
- The described chromatographic method is effective for high-scale purification of prostatic acid phosphatase (PAP).
- This purification strategy provides a foundation for detailed structural and functional analyses of PAP.
- The determination of PAP's 3D structure opens new avenues for understanding its biological roles and potential therapeutic targeting.
