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Journal Articles

A Methodology for estimating localization of apurinic/apyrimidinic sites in DNA using fluorescence resonance energy transfer

Akamatsu, Ken; Shikazono, Naoya

Analytical Biochemistry, 433(2), p.171 - 180, 2013/02

 Times Cited Count:5 Percentile:17.98(Biochemical Research Methods)

Journal Articles

Reconstitution of the muscle thin filament from recombinant troponin components and the native thin filaments

Matsumoto, Fumiko; Deshimaru, Shungo*; Oda, Toshiro*; Fujiwara, Satoru

Analytical Biochemistry, 399(2), p.299 - 301, 2010/04

 Times Cited Count:2 Percentile:58.78(Biochemical Research Methods)

We have developed a technique by which muscle thin filaments are reconstituted from the recombinant troponin components and the native thin filaments. By this technique, the reconstituted troponin complex is exchanged into the native thin filaments in the presence of 20% glycerol and 0.3 M KCl at pH 6.2. Over 90% of endogenous troponin complex was replaced with the recombinant troponin complex. Structural integrity and Ca$$^{2+}$$-sensitivity of the reconstituted thin filament prepared by this technique was confirmed by X-ray fiber diffraction measurements and the thin filament activated myosin subfragment 1 ATPase measurements, respectively.

Journal Articles

A Novel methodology for characterizing strand-break termini and damaged bases in plasmid DNA exposed to ionizing radiation

Akamatsu, Ken

Analytical Biochemistry, 362(2), p.229 - 235, 2007/03

 Times Cited Count:4 Percentile:13.53(Biochemical Research Methods)

We have developed a de novo methodology to characterize radiation damage in DNA. An enzyme system consisting of snake venom phosphodiesterase (SVPD) and calf intestine alkaline phosphatase (CIAP) was used to examine the 3'termini of strand break sites. In this study, we hypothesized that the strand-break termini can be divided into two categories. The apparent radiation-chemical yield (G-value) can be estimated using the level of intact 2'-deoxynucleotides produced during a 15-min incubation with SVPD. The G-value for total strand breaks in dried DNA irradiated with $$^{60}$$Co $$gamma$$-rays was estimated to be 0.1 micromol/J. Moreover, the G-values of CIAP-dependent and CIAP-independent SVPD sites were estimated to be 0.078 and 0.024 micromol/J, respectively. These values suggest that 3'-phosphate termini are more likely to be produced than 3'termini without phosphate.

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