Why don't you do a blast search in the human proteome to see whether there is a
human ortholog to the bacterial protein you co-purified, and consider also
whether the interaction would make any sense physiologically, of course. It
might be that you have discovered something interesting!
Jacob
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Jacob Pearson Keller
Northwestern University
Medical Scientist Training Program
Dallos Laboratory
F. Searle 1-240
2240 Campus Drive
Evanston IL 60208
lab: 847.491.2438
cel: 773.608.9185
email: j-kell...@northwestern.edu
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On Jan 8, 2010, at 7:41 AM, Sivaraman Padavattan wrote:
Dear all,
I am trying to express the human protein using bacterial expression strain
(Rosetta) and purified using Ni-NTA affinity purification. The Molecular weight
of out protein is 47 kDa. In SDS-PAGE, we have seen that 27 kDa contaminant
protein co-eluted with our protein even at high concentration of Imidazole. In
Superose 12 column, these two proteins eluted as a single peak and its
corresponding molecular weight suggestive of partial interaction. By mass
mapping we have found 27 kDa band is an adenylate kinase. Is there any specific
way to separate adenylate kinase from our protein?
Thanks in advance,
Sivaraman Padavattan
