caa a22 4500 445340851 CHVBK 20180317142812.0 cr unu---uuuuu 170323e20111001xx s 000 0 eng 10.1007/s00436-011-2347-y doi (NATIONALLICENCE)springer-10.1007/s00436-011-2347-y Cloning and characterization of a novel enzyme: tyrosine hydroxylase from Schistosoma japonicum [Elektronische Daten] [Yuansheng Hu, Dujuan Shi, Qingli Luo, Qingzhong Liu, Yindi Zhou, Lili Liu, Li Yu, Wei Wei, Jilong Shen] Catecholamines, such as dopamine and noradrenaline, play important roles as neuromuscular transmitters and modulators in all parasitic helminthes, including Schistosoma japonicum. S. japonicum tyrosine hydroxylase (SjTH) was amplified by rapid amplification of cDNA ends polymerase chain reaction that shows strong homology to Schistosoma mansoni tyrosine hydroxylase, the enzyme that catalyzes the first and rate-limiting step in the biosynthesis of catecholamines. The SjTH transcripts encoded the protein of 463 amino acids and a predicted size of 54kDa. Purified recombinant SjTH as an N-terminal histidine fusion protein expressed in Escherichia coli showed catalytic activity that was confirmed with 3H tyrosine uptake. The purified enzyme was found to have the same absolute requirement for a tetrahydrobiopterin cofactor and similar sensitivity to be inhibited by high concentration of the substrate, tyrosine, as the mammalian enzyme. Also, purified SjTH showed characteristic inhibition by catecholamine products. The phosphorylated peptide from SjTH could interact with Sj14-3-3 signal protein. This evidence indicates that SjTH encodes a functional tyrosine hydroxylase that has catalytic properties similar to those of the mammalian hosts' enzyme, and its catalytic activity could be regulated by a phosphorylated or dephosphorylated form. This demonstration of SjTH further suggests that the parasites have the enzymatic capacity to synthesize catecholamines endogenously. Springer-Verlag, 2011 Hu Yuansheng Institute of Clinical Pharmacology, Anhui Medical University, The Key Laboratories of Zoonoses and Pathogen Biology, Anhui, 230032, Hefei, Anhui Province, China aut Shi Dujuan Anhui Key Laboratories of Zoonoses and Pathogen Biology, 230032, Hefei, Anhui Province, China aut Luo Qingli Anhui Key Laboratories of Zoonoses and Pathogen Biology, 230032, Hefei, Anhui Province, China aut Liu Qingzhong Department of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, 325000, Wenzhou, Zhejiang, China aut Zhou Yindi Department of Clinical Laboratory, The Third Affiliated Hospital of Anhui Medical University, 230061, Hefei, Anhui Province, China aut Liu Lili Anhui Key Laboratories of Zoonoses and Pathogen Biology, 230032, Hefei, Anhui Province, China aut Yu Li Anhui Key Laboratories of Zoonoses and Pathogen Biology, 230032, Hefei, Anhui Province, China aut Wei Wei Institute of Clinical Pharmacology, Anhui Medical University, The Key Laboratories of Zoonoses and Pathogen Biology, Anhui, 230032, Hefei, Anhui Province, China aut Shen Jilong Institute of Clinical Pharmacology, Anhui Medical University, The Key Laboratories of Zoonoses and Pathogen Biology, Anhui, 230032, Hefei, Anhui Province, China aut Parasitology Research Springer-Verlag 109/4(2011-10-01), 1065-1074 0932-0113 109:4<1065 2011 109 436 https://doi.org/10.1007/s00436-011-2347-y text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00436-011-2347-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Hu Yuansheng Institute of Clinical Pharmacology, Anhui Medical University, The Key Laboratories of Zoonoses and Pathogen Biology, Anhui, 230032, Hefei, Anhui Province, China aut NATIONALLICENCE 700 1- Shi Dujuan Anhui Key Laboratories of Zoonoses and Pathogen Biology, 230032, Hefei, Anhui Province, China aut NATIONALLICENCE 700 1- Luo Qingli Anhui Key Laboratories of Zoonoses and Pathogen Biology, 230032, Hefei, Anhui Province, China aut NATIONALLICENCE 700 1- Liu Qingzhong Department of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, 325000, Wenzhou, Zhejiang, China aut NATIONALLICENCE 700 1- Zhou Yindi Department of Clinical Laboratory, The Third Affiliated Hospital of Anhui Medical University, 230061, Hefei, Anhui Province, China aut NATIONALLICENCE 700 1- Liu Lili Anhui Key Laboratories of Zoonoses and Pathogen Biology, 230032, Hefei, Anhui Province, China aut NATIONALLICENCE 700 1- Yu Li Anhui Key Laboratories of Zoonoses and Pathogen Biology, 230032, Hefei, Anhui Province, China aut NATIONALLICENCE 700 1- Wei Wei Institute of Clinical Pharmacology, Anhui Medical University, The Key Laboratories of Zoonoses and Pathogen Biology, Anhui, 230032, Hefei, Anhui Province, China aut NATIONALLICENCE 700 1- Shen Jilong Institute of Clinical Pharmacology, Anhui Medical University, The Key Laboratories of Zoonoses and Pathogen Biology, Anhui, 230032, Hefei, Anhui Province, China aut NATIONALLICENCE 773 0- Parasitology Research Springer-Verlag 109/4(2011-10-01), 1065-1074 0932-0113 109:4<1065 2011 109 436 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer