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Purification and characterization of mouse brain Thy-1.2 differentiation alloantigen
Brain Research
|December 22, 1978
Summary
Researchers purified Thy-1.2, a brain antigen, from mouse brains using subcellular fractionation and chromatography. The purified glycoprotein showed similarities to Thy-1.1, suggesting subtle structural differences account for antigenic distinctions.
Area of Science:
- Neurobiology
- Biochemistry
- Immunology
Background:
- Thy-1 is a cell surface glycoprotein found in the brain.
- Thy-1.2 is an antigenic variant of Thy-1 present in mouse brains.
Purpose of the Study:
- To purify and characterize Thy-1.2 from C57BI/6J mouse brains.
- To compare the biochemical properties of mouse brain Thy-1.2 with other Thy-1 variants.
Main Methods:
- Subcellular fractionation to isolate synaptosomes.
- Deoxycholate extraction and delipidation for solubilization.
- Lectin affinity and gel filtration chromatography for purification.
- SDS-PAGE and amino acid/carbohydrate analysis for characterization.
Main Results:
- Thy-1.2 was purified over 2000-fold from mouse brain homogenate.
- Purified Thy-1.2 yielded a single band on SDS-PAGE with a molecular weight of 24,000.
- Amino acid composition was similar to mouse Thy-1.1; carbohydrate analysis showed similarity to rat Thy-1.1.
Conclusions:
- Mouse brain Thy-1.2 is a glycoprotein with properties similar to other Thy-1 variants.
- Antigenic differences between Thy-1.1 and Thy-1.2 are not readily detectable by compositional analysis.