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

doi:10.1093/nar/28.5.e14

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)

University of Dundee

Research output: Contribution to journal › Article › peer-review

32 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 language	English
Pages (from-to)	e4
Number of pages	1
Journal	Nucleic Acids Research
Volume	28
Issue number	5
DOIs	https://doi.org/10.1093/nar/28.5.e14
Publication status	Published - 1 Mar 2000

ASJC Scopus subject areas

Genetics

Access to Document

10.1093/nar/28.5.e14

Cite this

@article{d3b198ca59484335bb294fcc0825c268,

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

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.",

author = "Yue, \{Bai Gong\} and Paul Ajuh and G{\"o}ran Akusj{\"a}rvi and Lamond, \{Angus I.\} and Kreivi, \{Jan Peter\}",

year = "2000",

month = mar,

day = "1",

doi = "10.1093/nar/28.5.e14",

language = "English",

volume = "28",

pages = "e4",

journal = "Nucleic Acids Research",

issn = "0305-1048",

publisher = "Oxford University Press",

number = "5",

}

TY - JOUR

T1 - Functional coexpression of serine protein kinase SRPK1 and its substrate ASF/SF2 in Escherichia coli

AU - Yue, Bai Gong

AU - Ajuh, Paul

AU - Akusjärvi, Göran

AU - Lamond, Angus I.

AU - Kreivi, Jan Peter

PY - 2000/3/1

Y1 - 2000/3/1

N2 - 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.

AB - 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.

UR - https://www.scopus.com/pages/publications/0034146376

U2 - 10.1093/nar/28.5.e14

DO - 10.1093/nar/28.5.e14

M3 - Article

C2 - 10666475

AN - SCOPUS:0034146376

SN - 0305-1048

VL - 28

SP - e4

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 5

ER -

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

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this