Hi all,
Any of you got some experience on purifying (and crystallizing)
VIL-rich proteins (actually, more particularly isoleucine rich). Tried
to purify my protein, but majority (if not all) of the proteins reside
in the inclusion bodies. Induction is at low temperature, overnight
and tried doing urea treatment, but oddly the his-tagged protein
'loses' it's ability to bind to nickel column. When unbound protein of
interest ran in size exclusion column, it's eluting at void volume.
Also tried with working on a different Bl21 strain (Rosetta and RIL),
out of luck.
Ile (I) 20 12.0%
Val (V) 14 8.4%
Leu (L) 13 7.8%
Asn (N) 12 7.2%
Glu (E) 12 7.2%
Ser (S) 11 6.6%
Lys (K) 11 6.6%
Any suggestion would be nice, or point to direction where other
successfully purify and crystallize sort of similar protein. BLAST
does not give me a close homologue of the protein that I can work
with, so no information on possible domains.
Thanks
Allan
