Doug wrote:
>> I am looking for proteins with prosthetic groups which fluoresce in the visible
> to use in protein folding or enzyme kinetic studies using laser induced fluorescence
> at wavelengths longer than 450 nm. I am aware of the flavoenzymes. Are there
> any other proteins which have visible fluorescence that are stable in solution,
> are enzymes, or may lend them selfs well to reversible denaturation? Are there
> dyes which can be used to label proteins for these types of studies?
I am not sure how green fluorescent protein (GFP) reacts to
denaturation, that could certainly be tried. Many proteins contain
Trp-residues, but their excitation maximum is 280 nm, which would not
work with your laser. Several artificial environmentally sensitive
fluorescent probes are commercially available, they are linked to -SH,
-NH2 or -COOH groups in the protein. A good introduction into this topic
is the catalogue of Molecular Probes (http://www.probes.com). Note also
that there are other methods, like circular dichroism, which may be
employed for denaturation and renaturation studies.
