On resveratrol: Christ, are people still taking this?? It has now been demonstrated that Resveratrol does not activate sirtuins -- the effect is a laboratory artifact resulting from the fact that the molecular "tag" ("Fluor de Lys") that Sinclair's group has been using to detect the activation (deacetylation) of various enzymatic targets by sirtuins actually decreases the binding affinity of sirtuins to their target protein.
Jason Wood, co-author of Sinclair apparently, states on sci.life-extension that the interpretations regarding the Fluor de Lys incident were completely exaggerated:
http://groups.google...3f639b250a431b5
Well, i am a little late to the party, but a few comments on this
thread (google won't let me post a reply to the original):
"Fundamental laboratory error" is an overstatement, and claims that the
whole thing was made up are simply false. Nor is the work "faulty."
The test used was a well characterized assay for deacetylase activity,
and the interpretation of the results was consistent with the data
generated. In short, here's the scoop--the fluorescent moiety attached
to the peptide turned out to decrease the binding affinity of the
peptide for SIRT1. Resveratrol binds SIRT1 and then increases the
affinity for this lower-affinity substrate. This increase is not seen
in a fluorescent-less peptide, which binds much tighter. We have also
observed that the results of these assays can vary depending on the
concentration of substrate and NAD that are used. If the starting
concentration is too high, even with the fluorescent peptide, the
effect goes away. Resveratrol binding to SIRT1 is gaining acceptance in
the scientific literature. I would point you to an excellent recent
paper in JBC from John Denu's lab (available free on pubmed) examining
the biochemistry in more detail. They find similarly that activation in
the assay depends on the fluorescent moiety, but note they agree that
resveratrol _can_ bind SIRT1 and change substrate affinity. The real
question is how tight are the actual proteins binding? (under
investigation) If in vivo targets bind with a Kd more akin to the
labelled peptide, rather than the unlabelled, one could easily imagine
how the effect might be physiologically relevant.
As far as the lack of reproducibility of in vivo yeast data--that is
puzzling I agree, especially since the authors of the study are among
the world's leaders in performing yeast lifespans. I will only say at
least one other lab has repeated the yeast results, several labs have
repeated the worm results (with sir2 dependence), and flies have been
confirmed in at least two labs (with sir2 dependence).
So yes, it is not as straight forward as we had hoped. No, we didn't
make the whole thing up, and the sky is not falling.
Armchair scientists should be careful in drawing conclusions and
flinging accusations when they do not have first hand knowledge of
what's going on.
Sincerely,
Jason Wood
Note that the refence to "recent paper in JBC from John Denu's lab" is obviously Michael's [2]. He continues:
my vitriol was directed primarily at others who were harsher than you.
You raise valid questions. I agree with you that resveratrol seems to
have a lot of great properties, and a good many of the effects are
likely to have little to do with sirtuins.
With regard to conjugates--
you are entirely correct. These should be researched in the in vitro
assays. I have heard rumors that this is currently being done. One
thing to remember is the angle that we approached this problem from. We
are not pharmacologists, and are primarily interested in answering
questions about the basic biology of sirtuins and how they relate to
aging in our simple model systems from a genetic and biochemical
direction. Flies, worms, and yeast do not have livers. It seemed to
make sense to begin in vivo testing with the compound that came out of
the screen, which was the unconjugated resveratrol. Science in the lab
frequently does not develop in quite the orderly way media reports or
even finished published papers project. That having been said, results
using the conjugates you mention would certainly be exciting from the
point of view of (and perhaps more relevant to) human biology, which is
what we all ultimately care about. I, with you, look forward to seeing
more data.
As I said in the another thread, this starts to get pretty technical for me (I actually read [2] for like an hour), I'll wait until this is settled and enjoy my glass of red wine in the meantime.