TY - JOUR
T1 - Structural characterization of the RLCK family member BSK8
T2 - A pseudokinase with an unprecedented architecture
AU - Grütter, Christian
AU - Sreeramulu, Shivakumar
AU - Sessa, Guido
AU - Rauh, Daniel
N1 - Funding Information:
We thank Andrew McKinnon, Anja Michalczyk, and the Dortmund Protein Facility for cloning and expression of the BSK8 constructs. We thank Ingrid Vetter and the beam line scientists at X10SA for expert assistance during data collection. Kevan M. Shokat and Megan Riel-Mehan are thanked for helpful discussions and comments on the manuscript. G.S. was funded by the Binational Science Foundation ( BSF 2007091 ). This work was supported by the German Federal Ministry for Education and Research through the German National Genome Research Network-Plus (grant no. BMBF 01GS08104 to D.R.).
PY - 2013/11/15
Y1 - 2013/11/15
N2 - Brassinosteroid signaling kinases (BSKs) are plant-specific receptor-like cytoplasmic protein kinases involved in the brassinosteroid signaling pathway. Unlike common protein kinases, they possess a naturally occurring alanine residue at the "gatekeeper" position, as well as other sequence variations. How BSKs activate downstream proteins such as BSU1, as well as the structural consequences of their unusual sequential features, was unclear. We crystallized the catalytic domain of BSK8 and solved its structure by multiple-wavelength anomalous dispersion phasing methods to a resolution of 1.5 Å. In addition, a co-crystal structure of BSK8 with 5-adenylyl imidodiphosphate (AMP-PNP) revealed unusual conformational arrangements of the nucleotide phosphate groups and catalytic key motifs, typically not observed for active protein kinases. Sequential analysis and comparisons with known pseudokinase structures suggest that BSKs represent constitutively inactive protein kinases that regulate brassinosteroid signal transfer through an allosteric mechanism.
AB - Brassinosteroid signaling kinases (BSKs) are plant-specific receptor-like cytoplasmic protein kinases involved in the brassinosteroid signaling pathway. Unlike common protein kinases, they possess a naturally occurring alanine residue at the "gatekeeper" position, as well as other sequence variations. How BSKs activate downstream proteins such as BSU1, as well as the structural consequences of their unusual sequential features, was unclear. We crystallized the catalytic domain of BSK8 and solved its structure by multiple-wavelength anomalous dispersion phasing methods to a resolution of 1.5 Å. In addition, a co-crystal structure of BSK8 with 5-adenylyl imidodiphosphate (AMP-PNP) revealed unusual conformational arrangements of the nucleotide phosphate groups and catalytic key motifs, typically not observed for active protein kinases. Sequential analysis and comparisons with known pseudokinase structures suggest that BSKs represent constitutively inactive protein kinases that regulate brassinosteroid signal transfer through an allosteric mechanism.
KW - BR signaling kinase (BSK)
KW - X-ray crystallography
KW - brassinosteroid (BR) signaling
KW - pseudokinase
KW - receptor-like protein kinases
UR - http://www.scopus.com/inward/record.url?scp=84886724648&partnerID=8YFLogxK
U2 - 10.1016/j.jmb.2013.07.034
DO - 10.1016/j.jmb.2013.07.034
M3 - מאמר
AN - SCOPUS:84886724648
VL - 425
SP - 4455
EP - 4467
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
SN - 0022-2836
IS - 22
ER -