All - I was just in a discussion about TLS and one thing that came out
that I hadn't been aware of is that for the Biso restraints Refmac
restrains the difference between the 'residual' Bs, i.e. with the TLS
contributions subtracted, not the 'total' Bs.  Now it seems to me that
this isn't quite correct, because it's the total motion of the atoms
that matters, i.e. the total mean square along-bond displacements for
bonded atoms should be equal.  However, I can see that in practical
terms it won't make any significant difference provided appropriate
precautions are taken with the choice of TLS groups.

What I mean by this is that at least for domain-sized groups the
difference between the TLS contributions to the Bs for bonded atoms
within the *same* TLS group will be very small (but maybe not so small
for secondary-structure element or residue-sized groups), so in that
case the difference between the residual Bs for bonded atoms will be
essentially equal to the difference between total Bs and it won't matter
which you restrain.  However for bonded atoms in *different* TLS groups
this won't necessarily be true.

So it seems to me that the safe option is to choose TLS groups for
domains, SSE's etc, such that there are flexible 'linkers' separating
them that are not assigned to a TLS group, so that the domains can move
essentially independently.  One can still test whether linked domains do
actually behave as though the motion occurs by libration or torsion at a
single bond (i.e. a rigid linker) connecting the domains by comparing
the TLS results for the flexible and rigid linker cases.

Of course in many cases Nature already provides flexible linkers
connecting domains, and presumably the very reason they're there is to
allow some independent, but tethered, motion (this is no doubt a
much-simplified view of what's happening since non-bonded contacts will
also affect the motion and the purpose of the linker may well be to
constrain the motion in some specific way).

So I was wondering whether people do generally choose TLS groups with
this in mind, and indeed does the TLSMD server take this into account
when selecting TLS groups?

Cheers

-- Ian


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