I would like to phosphorylate a serine in my recombinant protein. Is there a good, reliable method that is generally accepted? I do not mind using an artificial phosphate mimic, especially a non-reversible one. My protein lacks cysteine residues so I can easily mutate the serine to cysteine.
You could mutate the serine to glutamate or aspartate, giving it a negative charge to mimic phosphorylation. This is often done.
Unnatural amino acid incorporation is probably the most direct method available and is usually worth the extra cloning that might be necessary. O-phospho-L-serine is commercially available and strains and plasmids are available from Addgene. Links to the 2011 Science paper describing the process and suppliers of the necesssary materials are listed below.
http://www.sigmaaldrich.com/catalog/product/sigma/p0878?lang=en®ion=US
http://www.sciencemag.org/content/333/6046/1151.full
https://www.addgene.org/browse/article/4968/
Hi everybody,
Have you ever try the system published in Science Steven Bown ?
regards,
Nico
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