Functional coexpression of serine protein kinase SRPK1 and its substrateASF/SF2 in Escherichia coli
2000 (English)In: Nucleic Acids Research, ISSN 0305-1048, E-ISSN 1362-4962, Vol. 28, no 5, e14- p.Article in journal (Refereed) Published
Mammalian proteins expressed in Escherichia coli are used in a variety of applications. A major drawback in producing eukaryotic proteins in E.coli is that the bacteria lack most eukaryotic post-translational modification systems, including serine/threonine protein kinase(s). Here we show that a eukaryotic protein can be phosphorylated in E.coli by simultaneous expression of a mammalian protein kinase and its substrate. We show that in bacteria expressing SRPK1, ASF/SF2 becomes phosphorylated to a degree resembling native ASF/SF2 present in interphase HeLa cell nuclei. The E.coli phosphorylated ASF/SF2 is functional in splicing and, contrary to the unphosphorylated protein, soluble under native conditions.
Place, publisher, year, edition, pages
2000. Vol. 28, no 5, e14- p.
Medical and Health Sciences
IdentifiersURN: urn:nbn:se:uu:diva-52396DOI: 10.1093/nar/28.5.e14PubMedID: 10666475OAI: oai:DiVA.org:uu-52396DiVA: diva2:80305