I want to express a protein with this information; single chain, tertiary structure around 500 aa, i dont have any information about the protein beside that.
- Is disulfide ligand and disulfide bond is the same thing on protein?
- if it's the same thing, is there any possibilities for the disulfide bond is formed even though the real protein doesn't have it if i express it on the E. coli origami strain?
I've looked into it, and it seems that a disulfide ligand is a disulfide bond in the context of a reaction (In the reaction 2-mercapthanol + disulfide bond, the disulfide bond is a disulfide ligand because it is a component of the reaction). If anyone has a better definition, I would be happy to be corrected.
For the second part of your question, the cytoplasm of E. coli is a reducing environment, so unless the protein is localized to the periplasm, it is unlikely that you will see spontaneous disulfide bonds in a standard E. coli strain. However, the Origami strain has mutations that facilitate the formation of disulfide bonds, so in your case, it's a possibility.
If you are trying to express your protein without disulfide bonds, you should consider using a different strain (BL-21 is a good choice). You can also try reducing the disulfide chemically with 2-mercapthanol or dithiothreitol if you think that the tertiary structure of your protein will not be disrupted.
If you're trying to encourage a new disulfide bond to be formed, the Origami strain will probably work, but only if your protein folds in such a way that the cysteines in your protein come into close proximity during folding.
Dear Jonathan Jonathan many thanks for your very interesting technical question. As an inorganic chemist I'm certainly not a proven specialist in biochemistry. However, it is clear that disulfide bond and disulfide ligand are two different pairs of shoes. First of all, a disulfide bond is an -S–S- moiety within any molecule, i.e. two sulfur atoms connected to each other by a single bond. In addition, each of the two sulfur atoms forms a single bond two two other organic moieties. Ligands in our field of research (inorganic chemistry) are organic molecules or anions coordinated to metal atoms. In biochemistry, the term ligand has a somewhat different meaning. According to the respective Wikipedia entry "a ligand In biochemistry and pharmacology means a substance that forms a complex with a biomolecule to serve a biological purpose."
https://en.wikipedia.org/wiki/Ligand_(biochemistry)
Thus, in summary, a disulfide bond is an -S–S- bond within a molecule, and a disulfide ligand is an organic molecules which contains a disulfide bond.
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