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

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

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / genetics*
  • Gene Expression*
  • Models, Molecular*
  • Plant Proteins* / biosynthesis
  • Plant Proteins* / chemistry
  • Plant Proteins* / genetics
  • Plant Proteins* / isolation & purification
  • Protein Domains
  • Protein Engineering*
  • Protein Kinases* / biosynthesis
  • Protein Kinases* / chemistry
  • Protein Kinases* / genetics
  • Protein Kinases* / isolation & purification
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification

Substances

  • Plant Proteins
  • Recombinant Proteins
  • Protein Kinases
  • S-receptor kinase