Efficient expression of SRK intracellular domain by a modeling-based protein engineering.
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.
['Murase|Kohji|K|', 'Hirano|Yoshinori|Y|', 'Takayama|Seiji|S|', 'Hakoshima|Toshio|T|']
D017360:Arabidopsis D015870:Gene Expression D008958:Models, Molecular D010940:Plant Proteins D000072417:Protein Domains D015202:Protein Engineering D011494:Protein Kinases D011994:Recombinant Proteins
Protein engineering Protein kinase Self-incompatibility Stable expression