Arginine kinase from Myzostoma cirriferum, a basal member of annelids.
We assembled a phosphagen kinase gene from the Expressed Sequence Tags database of Myzostoma cirriferum, a basal member of annelids. The assembled gene sequence was synthesized using an overlap extension polymerase chain reaction method and was expressed in Escherichia coli. The recombinant enzyme (355 residues) exhibited monomeric behavior on a gel filtration column and showed strong activity only for l-arginine. Thus, the enzyme was identified as arginine kinase (AK). The two-substrate kinetic parameters were obtained and compared with other AKs. Phylogenetic analysis of amino acid sequences of phosphagen kinases indicated that the Myzostoma AK gene lineage differed from that of the polychaete Sabellastarte spectabilis AK, which is a dimer of creatine kinase (CK) origin. It is likely that the Myzostoma AK gene lineage was lost at an early stage of annelid evolution and that Sabellastarte AK evolved secondarily from the CK gene. This work contributes to our understanding of the evolution of phosphagen kinases of annelids with marked diversity.
Laboratory of Biochemistry, Faculty of Science, Kochi University, Kochi 780-8520, Japan.
Daichi
D
Yano
Laboratory of Biochemistry, Faculty of Science, Kochi University, Kochi 780-8520, Japan.
Sayo
S
Mimura
Laboratory of Biochemistry, Faculty of Science, Kochi University, Kochi 780-8520, Japan.
Kouji
K
Uda
Laboratory of Biochemistry, Faculty of Science, Kochi University, Kochi 780-8520, Japan. Electronic address: suzuki@kochi-u.ac.jp.
Tomohiko
T
Suzuki
D000595:Amino Acid Sequence; D000818:Animals; D000845:Annelida / Q000201:enzymology* / Q000235:genetics; D001122:Arginine Kinase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D020224:Expressed Sequence Tags / Q000378:metabolism; D007700:Kinetics; D016415:Sequence Alignment; D013045:Species Specificity
D001122:Arginine Kinase
Annelids; Arginine kinase; Creatine kinase; Myzostoma cirriferum; Phosphagen kinase