title: The Arabidopsis thaliana SERK1 kinase domain spontaneously refolds to an active state in vitro.
abstract: Auto-phosphorylating kinase activity of plant leucine-rich-repeat receptor-like kinases (LRR-RLK's) needs to be under tight negative control to avoid unscheduled activation. One way to achieve this would be to keep these kinase domains as intrinsically disordered protein (IDP) during synthesis and transport to its final location. Subsequent folding, which may depend on chaperone activity or presence of interaction partners, is then required for full activation of the kinase domain. Bacterially produced SERK1 kinase domain was previously shown to be an active Ser/Thr kinase. SERK1 is predicted to contain a disordered region in kinase domains X and XI. Here, we show that loss of structure of the SERK1 kinase domain during unfolding is intimately linked to loss of activity. Phosphorylation of the SERK1 kinase domain neither changes its structure nor its stability. Unfolded SERK1 kinase has no autophosphorylation activity and upon removal of denaturant about one half of the protein population spontaneously refolds to an active protein in vitro. Thus, neither chaperones nor interaction partners are required during folding of this protein to its catalytically active state.
authors.affiliation: Laboratory of Biochemistry, Wageningen University, Wageningen, The Netherlands. marije.aandentoorn@wur.nl
authors.forename: Marije
authors.initials: M
authors.lastname: Aan den Toorn
authors.forename: Mieke M E
authors.initials: MM
authors.lastname: Huijbers
authors.forename: Sacco C
authors.initials: SC
authors.lastname: de Vries
authors.forename: Carlo P M
authors.initials: CP
authors.lastname: van Mierlo
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D010766:Phosphorylation; D011487:Protein Conformation; D017510:Protein Folding; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*
chemical_list: D029681:Arabidopsis Proteins; D011494:Protein Kinases; C440035:SERK1 protein, Arabidopsis