Hypotaurocyamine kinase evolved from a gene for arginine kinase.
Hypotaurocyamine kinase (HTK) is a member of the highly conserved family of phosphagen kinases that includes creatine kinase (CK) and arginine kinase (AK). HTK is found only in sipunculid worms, and it shows activities for both the substrates hypotaurocyamine and taurocyamine. Determining how HTK evolved in sipunculids is particularly insightful because all sipunculid-allied animals have AK and only some sipunculids have HTK. We determined the cDNA sequence of HTK from the sipunculid worm Siphonosoma cumanense for the first time, cloned it in pMAL plasmid and expressed it in E. coli as a fusion protein with maltose-binding protein. The cDNAderived amino acid sequence of Siphonosoma HTK showed high amino acid identity with molluscan AKs. Nevertheless, the recombinant enzyme of Siphonosoma HTK showed no activity for the substrate arginine, but showed activity for taurocyamine. Comparison of the amino acid sequences of HTK and AK indicated that the amino acid residues necessary for the binding of the substrate arginine in AK have been completely lost in Siphonosoma HTK sequence. The phylogenetic analysis indicated that the HTK amino acid sequence was placed just outside the molluscan AK cluster, which formed a sister group with the arthropod and nematode AKs. These results suggest that Siphonosoma HTK evolved from a gene for molluscan AK. Moreover, to confirm this assertion, we determined by PCR that the gene for Siphonosoma HTK has a 5-exon/4-intron structure, which is homologous with that of the molluscan AK genes. Further, the positions of splice junctions were conserved exactly between the two genes. Thus, we conclude that Siphonosoma HTK has evolved from a primordial gene for molluscan AK.
Uda
Kouji
K
Laboratory of Biochemistry, Faculty of Science, Kochi University, Akebonocho 2-5-1, Kochi 780-8520, Japan.
Iwai
Aki
A
Suzuki
Tomohiko
T
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001122:Arginine Kinase / Q000737:chemistry / Q000235:genetics*; D002352:Carrier Proteins / Q000378:metabolism; D003001:Cloning, Molecular; D016000:Cluster Analysis; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D019143:Evolution, Molecular; D005091:Exons; D017238:Genes, Helminth; D006639:Histidine / Q000378:metabolism; D007438:Introns; D007532:Isoleucine / Q000378:metabolism; D007930:Leucine / Q000378:metabolism; D058619:Maltose-Binding Proteins; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D008970:Molecular Weight; D008974:Mollusca / Q000235:genetics; D009348:Nematoda / Q000201:enzymology / Q000235:genetics*; D016366:Open Reading Frames; D017852:Phosphotransferases (Nitrogenous Group Acceptor) / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
D002352:Carrier Proteins; D018076:DNA, Complementary; D058619:Maltose-Binding Proteins; D011993:Recombinant Fusion Proteins; D007532:Isoleucine; D006639:Histidine; D017852:Phosphotransferases (Nitrogenous Group Acceptor); D001122:Arginine Kinase; C507222:hypotaurocyamine kinase; D007930:Leucine