Related Experiment Videos
Characterisation of phosphoprotein complexes associated with rat thymocyte Thy-1
D Garnett1, L L Spruyt, A D Beyers
1Department of Medical Biochemistry, University of Stellenbosch, Tygerberg, South Africa.
Summary
Researchers found that the phosphoprotein p85, associated with rat thymocyte Thy-1, is an integral membrane protein. This protein is linked via disulfide bonds, indicating an extracellular domain and suggesting phosphatidylinositol 3 kinase activity association.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Protein tyrosine kinases p56lck and p60fyn associate with rat thymocyte Thy-1.
- Thy-1 is a cell surface glycoprotein involved in T-cell activation.
Purpose of the Study:
- To investigate the nature of phosphoproteins associated with rat thymocyte Thy-1.
- To determine if phosphatidylinositol 3 kinase activity is associated with Thy-1.
Main Methods:
- Disulfide bridging analysis to identify protein associations.
- Coprecipitation assays using Brij 96 thymocyte lysates.
Main Results:
- Phosphoprotein p85, associated with Thy-1, forms disulfide bridges with another polypeptide.
- This indicates p85 is an integral membrane protein with an extracellular domain.
- Phosphatidylinositol 3 kinase activity was coprecipitated with Thy-1.
Conclusions:
- Rat thymocyte Thy-1 is associated with an integral membrane phosphoprotein (p85) with an extracellular domain.
- Phosphatidylinositol 3 kinase activity is linked to Thy-1 in thymocyte lysates.