New structural insights and molecular-modelling studies of 4-methyl-5-beta-hydroxyethylthiazole kinase from Pyrococcus horikoshii OT3 (PhThiK).
4-Methyl-5-beta-hydroxyethylthiazole kinase (ThiK) catalyses the phosphorylation of the hydroxyl group of 4-methyl-5-beta-hydroxyethylthiazole. This work reports the first crystal structure of an archaeal ThiK: that from Pyrococcus horikoshii OT3 (PhThiK) at 1.85 A resolution with a phosphate ion occupying the position of the beta-phosphate of the nucleotide. The topology of this enzyme shows the typical ribokinase fold of an alpha/beta protein. The overall structure of PhThiK is similar to those of Bacillus subtilis ThiK (BsThiK) and Enterococcus faecalis V583 ThiK (EfThiK). Sequence analysis of ThiK enzymes from various sources indicated that three-quarters of the residues involved in interfacial regions are conserved. It also revealed that the amino-acid residues in the nucleotide-binding, magnesium ion-binding and substrate-binding sites are conserved. Binding of the nucleotide and substrate to the ThiK enzyme do not influence the quaternary association (trimer) as revealed by the crystal structure of PhThiK.
National Synchrotron Radiation Research Center, 101 Hsin-Ann Road, Hsinchu Science Park, Hsinchu 30076, Taiwan. kanthan@spring8.or.jp
Jeyaraman
J
Jeyakanthan
Subbiah
S
Thamotharan
Devadasan
D
Velmurugan
Vaijayanthimala Surya Narayna
VS
Rao
Shanthi
S
Nagarajan
Akeo
A
Shinkai
Seiki
S
Kuramitsu
Shigeyuki
S
Yokoyama
D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry; D003460:Crystallization; D018360:Crystallography, X-Ray; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry*; D020836:Protein Structure, Quaternary; D043902:Pyrococcus horikoshii / Q000201:enzymology*; D016415:Sequence Alignment
D019843:Archaeal Proteins; D017853:Phosphotransferases (Alcohol Group Acceptor); C051254:hydroxyethylthiazole kinase