Dear dpsk,

I asked a very similar question here about a month ago, so I'll post a
summary of the replies.

In our case, the protein was known to exist in a biologically relevant
monomer-dimer equilibrium, with the amount of monomer becoming
vanishingly small at high concentrations (e.g. in our crystallisation
assays). We obtained several crystal forms, one of which contained the
monomer and not the dimer.

Several people commented that the phenomenon isn't unusual - after all
what crystallises is not necessarily the predominant form of the
molecule but rather those species that have properties favouring a
crystalline phase in the given conditions. Upon nucleation, mass
action will replenish the pool of whatever crystallises at the expense
of the predominant species, so crystals can continue to grow even
though at any one time the concentration of the crystallising species
in solution is very low.

In spite of this, well-documented cases and/or systematic studies in
the literature seem to be fairly rare. Below is a list of replies with
relevant cases.

HTH,

best wishes,

Sebastiaan Werten.


Luca Jovine:

Sure - crystallization selects what packs, which may constitute a very
minor fraction of what you set up drops with. 1DUH is an example...


F.Xavier Gomis-Rüth:

we indeed found something very similar with selecase (see
http://www.ncbi.nlm.nih.gov/pubmed/25159620).
This is a highly specific metallopeptidase, which is also a
metamorphic protein that transits between several
stable states, among which only the monomer is the active species. We
managed to trap 5 different crystal
structures (monomers, three distinct dimers and tetramers)...and all
this in a small 110-residue protein!


Rick Lewis:

your crystallisation conditions could quite easily affect the MD
equilibrium. we had something similar a few years ago, when i was
working with a protein that was monomeric in solution 'til it was
phosphorylated when it then dimerised. in the crystals, however, we
had monomeric phospho-protein and (domain-swapped) dimeric
non-phosphoprylated. took some auc runs in conditions that mirrored
the xtl conditions to sort it out. have a look here:

http://www.ncbi.nlm.nih.gov/pubmed/11851334

i don't know of a more systematic study of multiple examples; i guess
people worry about their own problems first!

leo brady had something similar but different a few years ago with the
structure of a mis-folded cd2 variant, that was only a minor fraction
of the prep

http://www.ncbi.nlm.nih.gov/pubmed/7638192


Roger Rowlett:

some of the allosteric carbonic anhydrases stubbornly crystallize only
in the T-state, despite crystallization conditions that are known to
preferentially stabilize the R-state, and for which the predominant
R-state population can be confirmed by other methods.


Emilia C. Arturo (specifically commenting on proteins that not only
crystallise in different states but also change their
conformation/function depending on oligomerisation):

Our lab has termed a type of protein sequence that does this sort of
thing a "morpheein", and there is currently only one bonafide
morpheein in the family, though we are in the progress of validating
this term for at least one other enzyme. A description of morpheeins
can be found here: http://en.wikipedia.org/wiki/Morpheein



At Wednesday, 27-05-2015 on 04:56 dpsk wrote:


Hi,
Recentely,we determined a monomer protein structure which looks like an dimer 
in solution via size-exclusion chromatography and SAXS,
have you guys met such a situation before? or some references, 

Thank you for your reading

Best wishes,


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