Dear Firdous,
beryllium fluoride is actually a ground state analogue of GTP as
trifluoroberyllate is tetrahedral. To get a transition sate analogue you
need either AlF4- or MgF3-. Preparation of these complexes is very easy.
The great advantage of metal fluoride transition state analogue and
ground state analogue complexes is the fact that all components are
present in solution and readily self-assemble in the active site forming
stable enzyme complexes that are relevant to the catalytic cycle. The
inorganic metal fluoride salts (AlF3, and MgF2) are too insoluble to
use; therefore, the fluoride anion and metal cation components must be
added from separate stock solutions. Both ammonium fluoride and sodium
fluoride are suitable as the source of fluoride and are readily soluble
in water. Metal chlorides can be easily dissolved in water at high
concentration (0.5 M) and the solutions conserved at -20°C. One of the
critical aspects in preparing metal fluoride enzyme complexes is the pH
of the resulting solution. In particular, solutions of AlCl3 and BeCl2
are highly acidic (pH 2) samples should be prepared in 100 mM unbuffered
Tris base. The optimized sequence of component addition is to add
fluoride to the prepared buffer first, then the metal chloride stock.
Hope this helps, Matt
On 02/01/2024 18:53, Firdous Tarique wrote:
Hi
I would appreciate it if someone could share with me a step by step
protocol for making a stable GDP.BeF3 solution which is often used as
a transition state analogue for structural studies of a protein complex ?
Or a vendor from where it can be purchased directly.
Regards
Firdous
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Matthew Bowler
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European Molecular Biology Laboratory
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