title: Two arginine kinases of Tetrahymena pyriformis: characterization and localization.
abstract: Two cDNAs, one coding a typical 40-kDa arginine kinase (AK1) and the other coding a two-domain 80-kDa enzyme (AK2), were isolated from ciliate Tetrahymena pyriformis, and their recombinant enzymes were successfully expressed in Escherichia coli. Both enzymes had an activity comparable to those of typical invertebrate AKs. Interestingly, the amino acid sequence of T. pyriformis AK1, but not AK2, had a distinct myristoylation signal sequence at the N-terminus, suggesting that 40-kDa AK1 targets the membrane. Moreover, Western blot analysis showed that the AK1 is mainly localized in the ciliary fraction. Based on these results, we discuss the phosphoarginine shuttle, which enables a continuous energy flow to dynein for ciliary movement in T. pyriformis, and the role of AK1 in this model.
authors.affiliation: Laboratories of Biochemistry, Faculty of Science, Kochi University, Kochi 780-8520, Japan.
authors.forename: Juri
authors.initials: J
authors.lastname: Michibata
authors.affiliation: Laboratories of Biochemistry, Faculty of Science, Kochi University, Kochi 780-8520, Japan.
authors.forename: Noriko
authors.initials: N
authors.lastname: Okazaki
authors.affiliation: Laboratories of Biochemistry, Faculty of Science, Kochi University, Kochi 780-8520, Japan.
authors.forename: Shou
authors.initials: S
authors.lastname: Motomura
authors.affiliation: Laboratories of Biochemistry, Faculty of Science, Kochi University, Kochi 780-8520, Japan.
authors.forename: Kouji
authors.initials: K
authors.lastname: Uda
authors.affiliation: Cellular and Molecular Biotechnology, Faculty of Science, Kochi University, Kochi 780-8520, Japan.
authors.forename: Shigeki
authors.initials: S
authors.lastname: Fujiwara
authors.affiliation: Laboratories of Biochemistry, Faculty of Science, Kochi University, Kochi 780-8520, Japan. Electronic address: suzuki@kochi-u.ac.jp.
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Suzuki
mesh_terms: D000595:Amino Acid Sequence; D001122:Arginine Kinase / Q000235:genetics* / Q000378:metabolism; D020134:Catalytic Domain; D002923:Cilia / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D004398:Dyneins / Q000378:metabolism; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013769:Tetrahymena pyriformis / Q000201:enzymology*
chemical_list: D018076:DNA, Complementary; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D001122:Arginine Kinase; D004398:Dyneins
keywords: Arginine kinase; Myristoylation; Phosphoarginine shuttle; Subcellular localization; Tetrahymena pyriformis