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The murine interleukin 2 receptor. IV. Biochemical characterization
Journal of Immunology (Baltimore, Md. : 1950)
|June 1, 1986
Summary
Biochemical analysis of the Interleukin 2 (IL 2) receptor reveals a 32 kDa precursor protein. This precursor undergoes N-linked glycosylation and posttranslational modifications, resulting in a mature surface receptor with significant charge heterogeneity.
Area of Science:
- Immunology
- Molecular Biology
- Biochemistry
Background:
- The Interleukin 2 (IL 2) receptor plays a crucial role in immune responses.
- Understanding its structure and posttranslational modifications is key to deciphering its function.
Purpose of the Study:
- To biochemically characterize the IL 2 receptor from the CTL-L cell line.
- To investigate the precursor forms and posttranslational modifications of the IL 2 receptor.
Main Methods:
- Pulse-chase and tunicamycin studies.
- Endoglycosidase digestion (Endo-F, Endo-H).
- Two-dimensional gel electrophoresis (IEF vs SDS-PAGE).
- Western blot analysis using anti-IL 2 receptor monoclonal antibody (7D4).
Main Results:
- A single protein precursor (p32) was identified, rapidly processed into intermediates (p38, p40) via N-linked glycosylation.
- Mature surface receptor (p58) exhibited apparent molecular weights from 52,000 to 65,000 daltons and broad charge heterogeneity.
- An additional 110 kDa species (p110) was detected, sharing an identical isoelectric point with p58 and expressing the same epitope.
- Similar molecular weight bands were observed in mitogen-stimulated T and B lymphocytes, though slightly smaller.
Conclusions:
- The IL 2 receptor precursor undergoes significant N-linked glycosylation and other posttranslational modifications, contributing to its heterogeneity.
- The p110 species is likely associated with the IL 2 receptor rather than being a distinct molecule.
- Posttranslational modifications of the IL 2 receptor may vary between cell types.