Functional coexpression of serine protein kinase SRPK1 and its substrate ASF/SF2 in Escherichia coli

Bai Gong Yue, Paul Ajuh, Göran Akusjärvi, Angus I. Lamond, Jan Peter Kreivi (Lead / Corresponding author)

    Research output: Contribution to journalArticlepeer-review

    28 Citations (Scopus)

    Abstract

    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.

    Original languageEnglish
    Pages (from-to)e4
    Number of pages1
    JournalNucleic Acids Research
    Volume28
    Issue number5
    DOIs
    Publication statusPublished - 1 Mar 2000

    Fingerprint Dive into the research topics of 'Functional coexpression of serine protein kinase SRPK1 and its substrate ASF/SF2 in Escherichia coli'. Together they form a unique fingerprint.

    Cite this