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Complementary DNA for human T-cell cyclophilin
Insights
Researchers isolated and sequenced complementary DNA for human cyclophilin, a cyclosporin A-binding protein, from T-cells. Human cyclophilin shows minor differences from bovine cyclophilin and may belong to a multigene family.
Area of Science:
- Molecular Biology
- Immunology
- Genetics
Background:
- Cyclophilin is a specific cyclosporin A-binding protein.
- Understanding human cyclophilin is crucial for immunology and drug development.
Purpose of the Study:
- To isolate and sequence complementary DNA (cDNA) encoding human cyclophilin.
- To compare human cyclophilin with its bovine counterpart.
- To investigate the expression and gene family of human cyclophilin.
Main Methods:
- Complementary DNA isolation and sequencing from Jurkat T-cell line.
- Amino acid sequence comparison between human and bovine cyclophilin.
- RNA transfer blot analysis for mRNA size determination.
- Southern blot analysis for gene family investigation.
Main Results:
- Human cyclophilin cDNA was successfully isolated and sequenced from T-cells.
- Human cyclophilin exhibits high similarity to bovine cyclophilin, with only three amino acid differences.
- Cyclophilin mRNA levels are only marginally affected by T-cell induction and cyclosporin A treatment.
- Southern blot analysis suggests cyclophilin is encoded by a multigene family in humans.
Conclusions:
- Human cyclophilin is structurally similar to bovine cyclophilin.
- The expression of cyclophilin mRNA is relatively stable under specific induction conditions.
- The human genome likely contains multiple genes for cyclophilin, indicating functional diversity.
Abstract:
Complementary DNA encoding human cyclophilin, a specific cyclosporin A-binding protein, has been isolated from the leukemic T-cell line Jurkat and sequenced. Comparison of the deduced amino acid sequence with the previously determined sequence of bovine thymus cyclophilin reveals only three differences: an additional amino acid at the carboxy terminus end and two internal changes. RNA transfer blot analysis indicates an mRNA size of approximately 1 kb for human T-cell cyclophilin. Phytohaemagglutinin and phorbol myristate acetate induction of T cells treated or not with cyclosporin A affects only marginally the level of cyclophilin mRNA. Southern blot analysis of human genomic DNA digested with different restriction enzymes strongly suggests the existence of a multigene family for cyclophilin.