TY  - JOUR
AU  - Shibazaki, Chie
AU  - Arai, Shigeki
AU  - Shimizu, Rumi
AU  - Saeki, Morihisa
AU  - Kinoshita, Takayoshi
AU  - Ostermann, Andreas
AU  - Schrader, Tobias E.
AU  - Kurosaki, Yuzuru
AU  - Sunami, Tomoko
AU  - Kuroki, Ryota
AU  - Adachi, Motoyasu
TI  - Hydration Structures of the Human Protein Kinase CK2α Clarified by Joint Neutron and X-ray Crystallography
JO  - Journal of molecular biology
VL  - 430
IS  - 24
SN  - 0022-2836
CY  - Amsterdam [u.a.]
PB  - Elsevier
M1  - FZJ-2019-00781
SP  - 5094 - 5104
PY  - 2018
AB  - Casein kinase 2 (CK2) has broad phosphorylation activity against various regulatory proteins, which are important survival factors in eukaryotic cells. To clarify the hydration structure and catalytic mechanism of CK2, we determined the crystal structure of the alpha subunit of human CK2 containing hydrogen and deuterium atoms using joint neutron (1.9 Å resolution) and X-ray (1.1 Å resolution) crystallography. The analysis revealed the structure of conserved water molecules at the active site and a long potential hydrogen bonding network originating from the catalytic Asp156 that is well known to enhance the nucleophilicity of the substrate OH group to the γ-phospho group of ATP by proton elimination. His148 and Asp214 conserved in the protein kinase family are located in the middle of the network. The water molecule forming a hydrogen bond with Asp214 appears to be deformed. In addition, mutational analysis of His148 in CK2 showed significant reductions by 40%–75% in the catalytic efficiency with similar affinity for ATP. Likewise, remarkable reductions to less than 5% were shown by corresponding mutations on His131 in death-associated protein kinase 1, which belongs to a group different from that of CK2. These findings shed new light on the catalytic mechanism of protein kinases in which the hydrogen bond network through the C-terminal domain may assist the general base catalyst to extract a proton with a link to the bulk solvent via intermediates of a pair of residues.
LB  - PUB:(DE-HGF)16
C6  - pmid:30359582
UR  - <Go to ISI:>//WOS:000454377100019
DO  - DOI:10.1016/j.jmb.2018.09.018
UR  - https://juser.fz-juelich.de/record/859975
ER  -