Related Experiment Video
Updated: Jul 16, 2026
10:46
Production of Human Norovirus Protruding Domains in E. coli for X-ray Crystallography
Published on: April 19, 2016
Expression, purification and crystallization of human CD5 domain III, a nano-scale crystallization example
Bernardo Rodamilans1, Sonia Ibañez, Elisabeth Bragado-Nilsson
1Structural Biology and Biocomputing Programme, Macromolecular Crystallography Group, Spanish National Cancer Center (CNIO) c/Melchor Fdez. Almagro 3, 28029 Madrid, Spain.
Journal of Structural Biology
|March 6, 2007
Summary
Structural insights into CD5 domain III, a key immune regulator, were obtained. This study presents a method for producing and crystallizing difficult-to-express mammalian proteins for structural analysis.
Area of Science:
- Immunology
- Structural Biology
- Biochemistry
Background:
- CD5 is a crucial regulator of T cell activation and immune responses.
- CD5 is a membrane glycoprotein in the scavenger receptor cysteine-rich (SRCR) superfamily.
- No prior structural information was available for CD5.
Purpose of the Study:
- To obtain structural information of the CD5 protein.
- To develop a method for producing and crystallizing CD5 domain III.
- To enable structural studies of challenging mammalian proteins.
Main Methods:
- Expressed the conserved membrane-proximal SRCR domain of CD5 (domain III) in HEK-EBNA-293 cells.
- Purified the expressed CD5 domain III protein.
- Obtained well-diffracting crystals of CD5 domain III.
Main Results:
- Crystals belonged to a tetragonal space group (P4(1)22 or P4(3)22).
- Crystals diffracted to 2.5A resolution using synchrotron radiation.
- Two molecules were present per asymmetric unit.
Conclusions:
- The study successfully produced and crystallized CD5 domain III, yielding high-resolution diffraction data.
- The presented strategy can be applied to produce and crystallize other difficult-to-express mammalian proteins.
- This work paves the way for future structural and biophysical studies of CD5 and related proteins.
