title: Phosphagen kinase in Schistosoma japonicum: characterization of its enzymatic properties and determination of its gene structure.
abstract: Phosphagen kinases (PKs) play a major role in the regulation of energy metabolism in animals. Creatine kinase (CK) is the sole PK in vertebrates, whereas several PKs are present in invertebrates. Here, we report the enzymatic properties and gene structure of PK in the trematode Schistosoma japonicum (Sj). SjPK has a unique contiguous dimeric structure comprising domain 1 (D1) and domain 2 (D2). The three states of the recombinant SjPK (D1, D2, and D1D2) show a specific activity for the substrate taurocyamine. The comparison of the two domains of SjPK revealed that D1 had a high turnover rate (kcat=52.91) and D2 exhibited a high affinity for taurocyamine (Km(Tauro) =0.53±0.06). The full-length protein exhibited higher affinity for taurocyamine (Km(Tauro) =0.47±0.03) than the truncated domains (D1=1.30±0.10, D2=0.53±0.06). D1D2 also exhibited higher catalytic efficiency (kcat/Km(Tauro) =82.98) than D1 (40.70) and D2 (29.04). These results demonstrated that both domains of SjTKD1D2 interacted efficiently and remained functional. The three-dimensional structure of SjPKD1 was constructed by the homology modeling based on the transition state analog complex state of Limulus AK. This protein model of SjPKD1 suggests that the overall structure is almost conserve between SjPKD1 and Limulus AK except for the flexible loops, that is, particularly guanidino-specificity (GS) region, which is associated with the recognition of the corresponding guanidino substrate. The constructed NJ tree and the comparison of exon/intron organization suggest that SjTK has evolved from an arginine kinase (AK) gene. SjTK has potential as a novel antihelminthic drug target as it is absent in mammals and its strong activity may imply a significant role for this protein in the energy metabolism of the parasite.
authors.affiliation: Department of Environmental Health Sciences, Kochi Medical School, Oko, Nankoku City, Kochi 783-8505, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Tokuhiro
authors.forename: Kouji
authors.initials: K
authors.lastname: Uda
authors.forename: Hiroko
authors.initials: H
authors.lastname: Yano
authors.forename: Mitsuru
authors.initials: M
authors.lastname: Nagataki
authors.forename: Blanca R
authors.initials: BR
authors.lastname: Jarilla
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Suzuki
authors.forename: Takeshi
authors.initials: T
authors.lastname: Agatsuma
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016000:Cluster Analysis; D018509:DNA, Helminth / Q000737:chemistry / Q000235:genetics; D019143:Evolution, Molecular; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017852:Phosphotransferases (Nitrogenous Group Acceptor) / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D010802:Phylogeny; D011487:Protein Conformation; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D012549:Schistosoma japonicum / Q000201:enzymology* / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013654:Taurine / Q000031:analogs & derivatives / Q000378:metabolism
chemical_list: D018509:DNA, Helminth; D013654:Taurine; C009558:taurocyamine; D017852:Phosphotransferases (Nitrogenous Group Acceptor); C022716:taurocyamine kinase