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Updated: Jul 11, 2026
Mutagenesis and Analysis of Genetic Mutations in the GC-rich KISS1 Receptor Sequence Identified in Humans with Reproductive Disorders
Published on: September 4, 2011
Molecular cloning and overexpression of the human FK506-binding protein FKBP
R F Standaert1, A Galat, G L Verdine
1Department of Chemistry, Harvard University, Cambridge, Massachusetts 02138.
Abstract:
The potent immunosuppressive agent FK506 is highly effective in preventing organ transplant rejection in humans. Like cyclosporin A, FK506 inhibits the transcription of early T-cell activation genes, apparently by modulating the activity of transcriptional regulators such as nuclear factor of activated T cells. A remarkable finding is that the predominant binding proteins (immunophilins) for cyclosporin A and FK506, cyclophilin and FKBP respectively, are peptidyl-prolyl-cis-trans-isomerases that are potently and selectively inhibited by their respective ligands. Here we report the complementary DNA and derived amino-acid sequences of human FKBP from Jurkat cells and also the efficient overexpression in Escherichia coli of fully active, recombinant human FKBP. The human FKBP cDNA sequence shows significant similarity to an open reading frame in the Neisseria meningitidis genome.
Insights
The immunosuppressive drug FK506 binds to FKBP, a peptidyl-prolyl isomerase. Researchers sequenced human FKBP and expressed it in E. coli, finding similarity to a Neisseria meningitidis gene.
Area of Science:
- Immunology
- Molecular Biology
- Biochemistry
Background:
- FK506 is a potent immunosuppressant crucial for preventing organ transplant rejection.
- FK506, like cyclosporin A, inhibits T-cell activation gene transcription by modulating transcription factors.
- FK506 binds to FKBP, a peptidyl-prolyl cis-trans isomerase, which is potently inhibited by FK506.
Purpose of the Study:
- To determine the complementary DNA (cDNA) and amino acid sequences of human FKBP.
- To achieve efficient overexpression of active, recombinant human FKBP in Escherichia coli.
- To investigate potential evolutionary or functional links by comparing human FKBP to other genomes.
Main Methods:
- Isolation and sequencing of complementary DNA (cDNA) encoding human FKBP from Jurkat cells.
- Cloning and expression of recombinant human FKBP in Escherichia coli for overexpression.
- Bioinformatic analysis to compare the human FKBP sequence with existing genomic databases.
Main Results:
- The cDNA and derived amino acid sequences of human FKBP were successfully determined.
- Active, recombinant human FKBP was efficiently overexpressed in Escherichia coli.
- The human FKBP cDNA sequence exhibited significant similarity to an open reading frame in the Neisseria meningitidis genome.
Conclusions:
- Human FKBP, a key binding protein for the immunosuppressant FK506, has been characterized at the sequence level.
- The successful overexpression of recombinant human FKBP facilitates further biochemical and structural studies.
- The sequence similarity suggests a conserved function or evolutionary origin of FKBP across different species, including bacteria.
