Dear all,
Sorry for the slightly off-topic question.
I am working on a non-native, *de novo* [4Fe-4S]
protein, designed as a four-helix bundle. The *in vitro* reconstituted
protein assembles with [4Fe-4S] (confirmed by EPR) and exists in
monomer-dimer configuration (confirmed by SEC). These results have been
already published.
Recently we could get the [4Fe-4S] assembly directly from
the *E. coli* (*in vivo* assembly). Everything is as expected (compared to
reconstituted protein), except the oligomerization state. The protein
assembles as trimer, in contrast to monomer-dimer configuration of the
reconstituted protein. The trimeric nature of the *in vivo* assembled
protein has been confirmed by SEC, SEC-SLS and SAXS.
*So, my question is, has anyone encountered such situation,
where the As-purified Fe-S protein having a completely different oligomeric
state compared to the in vitro reconstitution protein? *
Looking forward to hearing for some examples and/or references.
Regards,
Bhanu
