Abstract
Genetic analyses have revealed that leucine-rich repeat receptor-like kinases (LRR-RLKs) play important roles in plant signal transduction. However, biochemical properties of most LRR-RLKs remain unknown because it is difficult to obtain sufficient proteins for biochemical analyses. Therefore, we optimized LRR-RLK expression in Saccharomyces cerevisiae.
First, to determine the most appropriate strain for LRR-RLK expression, LRR-RLKs we expressed in several strains and found that BY2777, which is a mutant strain of vacuolar proteases is appropriate strain for it. In yeast, it is known that proper folding and secretion of some protein can be facilitated by fusing it to yeast-derived polypeptide Hsp150Δ. Therefore, we also examined whether Hsp150Δcan promote the secretion of LRR-RLK in yeast. Secretions of some LRR-RLKs are found to be improved by fusing them to Hsp150Δ. We are now optimizing LRR-RLK solubilization conditions and confirming their kinase activity for the high-throughput assessment of protein quality.
