7 research outputs found
Electronically preresonant stimulated Raman scattering microscopy in the visible
We report an experimental scheme for stimulated Raman scattering (SRS) microscopy with excitation in the visible spectral region. This allows electronically preresonant (epr) SRS microscopy of a broad range of chromophores with sensitivities as low as 1 μM. Our experiment is based on two synchronously near-infrared pumped optical parametric oscillators (OPO). One of the outputs is modulated at a fourth of the repetition rate with a novel broadband electro-optical modulator. Using a combination of spectral focusing and tuning of the OPO, we show the recording of epr-SRS spectra over the whole range of molecular vibrations at a speed up to 20 times faster than classical wavelength tuning. The imaging capabilities of this setup are demonstrated with material scientific and cellular samples
The prolyl isomerase, FKBP25, interacts with RNA-engaged nucleolin and the pre-60S ribosomal subunit
FKBP (FK506 Binding Protein)
In the 70s, after a decade from the purification of cyclosporine, a selective immunosuppressant
agent and potent tool in transplantation medicine, a novel molecule was purified from bacteria
Streptomyces tsukubaensis. This molecule, called FK506, showed the same selective
immunosuppressant action as cyclosporine but was 10 to 100 fold more potent.
In an attempt to clarify the molecular mechanism through which the new drug exerted such a
selective effect on T-cells activation, two laboratories identified the cytosolic receptor for FK506.
This so-called FK506 binding protein (FKBP) was purified from bovine thymus, human spleen, and
Jurkat T-cell line. The isolated FKBP had an approximate molecular mass of 14 kDa and showed
an isomerase activity similar to the recently purified cyclosporine-binding protein, cyclophilin, but, it
was inhibited by FK506 and rapamycin but not cyclosporine. The
subsequent cloning of FKBP gene revealed that FKBP and cyclophilin had dissimilar sequences in
spite of their common enzymatic activity. The identified FKBP gene encoded for a protein of 108
aminoacids with a relative molecular mass of 11,819. For this reason, the progenitor of this
nascent class of proteins was later known as FKBP12.
The subsequent studies showed that FKBP12 was just a member of a ubiquitous and evolutionarily
conserved sub-family of proteins which differ from each other in their molecular weight and
structure. All FKBPs share a highly conserved domain, termed “FK-12 like domain”, capable of
binding to FK506 and exerting isomerase properties, i.e. interconversion from cis-to-trans and
trans-to-cis of peptide bonds involving proline, on protein substrates.
A schematic historical background of the 17 FKBPs so far identified is shown. A general
overview of FKBP structure, function and eventually associated disease is given in this
monograph, with the order of proteins following the chronology of discovery