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Protein Expr Purif. 2017 Mar;131:70-75. doi: 10.1016/j.pep.2015.09.020. Epub 2015 Sep 21.

Efficient expression of SRK intracellular domain by a modeling-based protein engineering.

Author information

1
Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan.
2
Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan. Electronic address: hakoshima@bs.naist.jp.

Abstract

S-locus protein kinase (SRK) is a receptor kinase that plays a critical role in self-recognition in the Brassicaceae self-incompatibility (SI) response. SRK is activated by binding of its ligand S-locus protein 11 (SP11) and subsequently induced phosphorylation of the intracellular kinase domain. However, a detailed activation mechanism of SRK is still largely unknown because of the difficulty in stably expressing SRK recombinant proteins. Here, we performed modeling-based protein engineering of the SRK kinase domain for stable expression in Escherichia coli. The engineered SRK intracellular domain was expressed about 54-fold higher production than wild type SRK, without loss of the kinase activity, suggesting it could be useful for further biochemical and structural studies.

KEYWORDS:

Protein engineering; Protein kinase; Self-incompatibility; Stable expression

PMID:
26390940
DOI:
10.1016/j.pep.2015.09.020
[Indexed for MEDLINE]

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