"Guy C. Brown" wrote:
>> We have not got arround to applying the analysis experimentally
> which would be nice. This would require an enzyme where the significant
> intermediates could be measured relatively easily in the steady state.
> Any ideas?
I can't think very quickly of an enzyme producing easily detectable
multiple intermediates on a multi-step reaction pathway. However, it
should be interesting even to observe the /equilibrium/ amounts of
reactants and products for a simple /reversible/ mechanism, presenting
itself (macroscopically) as
E + A <==> EX <==> E* + P <==> E + P
where E* is the "iso-enzyme".
I am thinking about some sort of an isomerase.
I suppose everybody remembers Knowles' et al. 1980s work on the
energetics of the "perfect enzyme". Those were relatively simple
enzymes (at least on the surface of things: neglecting oligomerization
of subunits, etc.) from which people derived surprisingly large amounts
of mechanistic information, to the extent that most of the elementary
rate constants are known quite accurately.
Maybe the horrendous formalism that Knowles at al. invented for their
"perfect enzyme" work could be subjected to yet another kind of torture,
and cast in terms of MCA?? I haven't thought this through in any
precise way, just brainstorming about enzymes where "significant
intermediates [in this case, products] could be measured relatively
easily in the steady state".
-- Petr
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Petr Kuzmic, Ph.D. * BioKin, Ltd. * Consulting & Software Development
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