Related Experiment Video
Updated: Jun 12, 2026
07:08
Efficient Mammalian Cell Expression and Single-step Purification of Extracellular Glycoproteins for Crystallization
Published on: December 23, 2015
Expression, purification and crystallization of human prolylcarboxypeptidase
Pravien D Abeywickrema1, Sangita B Patel, Noel J Byrne
1Global Structural Biology, Merck Research Laboratories, West Point, PA 19486, USA.
Summary
Prolylcarboxypeptidase (PrCP), an enzyme linked to hypertension and obesity, was successfully produced and crystallized. This breakthrough enables further structural studies of PrCP, crucial for understanding its role in disease.
Area of Science:
- Biochemistry
- Structural Biology
- Enzymology
Background:
- Prolylcarboxypeptidase (PrCP) is a lysosomal serine carboxypeptidase.
- PrCP cleaves C-terminal amino acids next to proline.
- PrCP is implicated in diseases like hypertension and obesity.
Purpose of the Study:
- To achieve robust production and purification of glycosylated human PrCP.
- To crystallize human PrCP for structural analysis.
- To facilitate further research into PrCP's role in disease.
Main Methods:
- Stable transformation of CHO cells for PrCP expression.
- Purification of glycosylated human PrCP.
- Crystallization of purified PrCP.
Main Results:
- Successful production and purification of glycosylated human PrCP.
- Obtained PrCP crystals in space group R32.
- Crystals diffracted to better than 2.8 A resolution.
Conclusions:
- The study reports the successful production, purification, and crystallization of human PrCP.
- The obtained crystal structure data will aid in understanding PrCP function.
- This work provides a foundation for structure-based drug design targeting PrCP.
