Based on my western blot result of a recombinant protein, there are two forms of the protein - soluble and insoluble. I was wondering, why is it important to acquire recombinant proteins in a soluble form? Does the insoluble form of protein means the failure of being folded to the right conformational shape?
Another question, is human interleukin 2 protein a form of soluble protein?
Jovan,
If your protein is naturally a soluble protein then finding it insoluble is usually because the expression system made it too fast or did not provide sufficient post-translational modification in order for your expressed protein to fold properly. It is of the utmost importance to get it to be soluble if it is meant to be a soluble protein. This will ensure that your expressed protein is adequately folded and that it is perform its normal function.
Hope this helps,
Marc
It is easier to purify protein as soluble form than as insoluble form such as inclusion bodies. As Jovan explained if a protein is expressed to fast it may not get sufficient post translational modification. To avoid it, grow the bacterial culture slower, for example at 30C, induce your recombinant protein much later and in shorter time, 1-2 hours.
That was helpful. However, can you recommend some journals that can explain this situation so that i can further understand the bigger picture? I can't seem to find a relevant journal to support this argument.
In general yes, insoluble protein = misfolded protein and viceversa. However beware that exceptions abound, as the actual picture is way more complex and there are cases of soluble misfolded aggregates, and of insoluble, correctly folded aggregates. See PMID 19046444, 16156893 and 15927061 for examples (there are many more in the literature).
Human IL-2 produced conventionally in E. coli does form aggregates. Purifying it from inclusion bodies is easy, though, as it is easy to refold.
Hope it helps!
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