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Kuwabara, Tomoko,Warashina, Masaki,Kato, Yoshio,Kawasaki, Hiroaki,Taira, Kazunari Korean Society for Biochemistry and Molecular Biol 2001 Journal of biochemistry and molecular biology Vol.34 No.1
Attempts using in vitro and in vivo selection procedures have been made to search for hammerhead ribozymes that have higher activities than the wild-type ribozyme and also to determine whether other sequences might be possible in the catalytic core of the hammerhead ribozyme. Active sequences selected in the past conformed broadly to the consensus core sequence except at A9, and no sequences were associated with higher activity than that of the hammerhead with the consensus core, an indication that the consensus sequence derived from viruses and virusoids is probably the optimal sequence [Vaish et al. (1997) Biochemistry 36, 6495-6501]. Recently, during construction of ribozyme expression vectors, we isolated a mutant hammerhead ribozyme, with an insertion of G between A9 and G10.1, that appeared to show significant activity [Kawasaki et al. (1996) Nucleic Acids Res. 24, 3010-3016; Kawasaki et al. (1998) Nature 393, 284-289]. We, therefore, characterized kinetic properties of the G-inserted mutant ribozymes in terms of the NUX rule. We demonstrate that the NUX rule is basically applicable to the G-inserted ribozymes and, more importantly, one type of G-inserted ribozyme was very active with $k_{cat}$, value of $6.4\;min^{-1}$ in 50 mM Tris-HCl (pH 8.0) and 10 mM $MgCl_2$ at $37^{\circ}C$.
Kuwabara, Tomoko,Warashina, Masaki,Taira, Kazunari,Kato, Yoshio,Kawasaki, Hiroaki 생화학분자생물학회 2002 BMB Reports Vol.34 No.1
Attempts using in vitro and in vivo selection procedures have been made to search for hammerhead ribozymes that have higher activities than the wild-type ribozyme and also to determine whether other sequences might be possible in the catalytic core of the hammerhead ribozyme. Active sequences selected in the past conformed broadly to the consensus core sequence except at A9, and no sequences were associated with higher activity than that of the hammerhead with the consensus core, an indication that the consensus sequence derived from viruses and virusoids is probably the optimal sequence [Vaish et al. (1997) Biochemistry 36, 6495-6501]. Recently, during construction of ribozyme expression vectors, we isolated a mutant hammerhead ribozyme, with an insertion of G between A9 and 610.1, that appeared to show significant activity [Kawasaki et at (1996) Nucleic Acids Res. 24, 3010-3016; Kawasaki et al. (1998) Nature 393, 284-289]. We, therefore, characterized kinetic properties of the G-inserted mutant ribozymes in terms of the NUX rule. We demonstrate that the NUX rule is basically applicable to the G-inserted ribozymes and, more importantly, one type of G-inserted ribozyme was very active with k_(cat), value of 6.4 min^(-1) in 50 mM Tris-HCl (pH 8.0) and 10 m1VI MgCl₂ at 37℃.
MAXIZYMEs: Allosterically controllable ribozymes with biosensor functions
Kurata, Hiroyuki,Miyagishi, Makoto,Kuwabara, Tomoko,Warashina, Masaki,Taira, Kazunari Korean Society for Biochemistry and Molecular Biol 2000 Journal of biochemistry and molecular biology Vol.33 No.5
Ribozymes are catalytic RNAs that can cleave RNAs at specific sites, thus they have been employed to degrade a target mRNA in vivo. Development of allosterically controllable ribozymes is of great current interest, but it remained difficult to furnish such functions to ribozymes in cultured cells or in animals. Recently, we designed allosterically controllable ribozymes termed maxizymes, which have sensor arms that recognize target mRNA sequences and, in the presence of such target sequences only, they form a cavity that can capture catalytically indispensable $Mg^{2+}$ ions, cleaving the target. The maxizyme was applied to therapy for chronic myelogenous leukemia (CML). It cleaved specifically the chimeric BCR-ABL mRNA, which caused CML, without damaging the normal ABL or BCR mRNA in mammalian cells and also in mice, providing the first successful example for allosteric control of the activity of artificial ribozymes in vivo.
Sonoko Mashimo,Naruto Yoshida,Ayaka Takegami,Yuki Warashina,Hitoshi Shiraki 대한운동학회 2019 아시아 운동학 학술지 Vol.21 No.2
[OBJECTIVES] To understand the condition of athletes engaged in daily sports activities, it is important to investigate the pain that many such athletes can experience as well as the pain that can be an initial symptom of injury. Although handball is a contact sport associated with frequent injuries, the actual nature of pain symptoms that develop has not been sufficiently studied, and the relationship between pain occurrence and athletes’ daily changing physical load remains unclear. This study aimed to clarify the nature of pain symptoms in handball players and examine the relationship between pain occurrence and physical load. [METHODS] This study involved a 12-month daily assessment of pain symptoms and physical load involving 11 university female handball players. Pain was examined in terms of body region and a pain severity score using a pain questionnaire, and physical load on handball was assessed based on playing hours, types of matches and training recorded by a video camera. [RESULTS] The total number of pains was 1698, and the pain incidence rate was 288.1 pains per 1000 player hours. In terms of body regions, the ankle (18.3%) was the most common, followed by the lower back (13.3%), foot (12.8%), Achilles tendon (9.2%), and thigh (8.9%). With regard to the relationship between pain and physical load, significant moderate or weak correlations were observed between handball (rs=0.657), training (rs=0.626), and on-court training (rs=0.591) and overall pain occurrences. In terms of body regions, the ankle and thigh, significant moderate or weak correlations were observed with respect to all categories in on-court training. [CONCLUSIONS] We found that pain occur frequently and athletes continue to compete in games despite experiencing pain. In addition, pain occurrence was related to physical load, indicating that the type of physical load depends on the body region.
MAXIZYMEs : Allosterically controllable ribozymes with biosensor functions
Kuratam Hiroyuki,Miyagishi, Makoto,Kuwabara, Tomoko,Warashina, Masaki,Taira, Kazunari 생화학분자생물학회 2001 BMB Reports Vol.33 No.5
Ribozymes are catalytic RNAs that can cleave RNAs at specific sites, thus they have been employed to degrade a target mRNA in vivo. Development of allosterically controllable ribozymes is of great current interest, but it remained difficult to furnish such functions to ribozymes in cultured cells or in animals. Recently, we designed allosterically controllable ribozymes termed maxizymes, which have sensor arms that recognize target mRNA sequences and, in the presence of such target sequences only, they form a cavity that can capture catalytically indispensable Mg^(2+) ions, cleaving the target. The maxizyme was applied to therapy for chronic myelogenous leukemia (CML). It cleaved specifically the chimeric BCR-ABL mRNA, which caused CML, without damaging the normal ABL or BCR mRNA in mammalian cells and also in mice, providing the first successful example for allosteric control of the activity of artificial ribozymes in vivo.