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Kaneko M, Masui R, Ake K, Kousumi Y, Kuramitsu S, Yamaguchi M, Kuyama H, Ando E, Norioka S, Nakazawa T, Okamura TA, Yamamoto H, Ueyama N: Rapid and sensitive amino-acid sequencing of cloning Thermus thermophilus HB8 ferredoxin by proteomics. J Proteome Res. 2004 Sep-Oct;3(5):983-7.
Recombinant holo Thermus thermophilus [7Fe-8S] ferredoxin was synthesized by cloning from Thermus thermophilus HB8 gene. A specific sequence (Pro-His-Val-Ile) at the N-terminus of the recombinant ferredoxin was determined by a rapid and highly sensitive mass spectral method using a novel Ru (II) Edman reagent, [(tpy) Ru (tpy-C6H4-NCS)](PF6) 2 (tpy=terpyridine). The formation of the recombinant holoTtFd was established by the characteristic absorptions and CD extrema as [7Fe-8S] ferredoxin. The catalytic electron-transfer reactivity of the [7Fe-8S] ferredoxin between ferredoxin-NADP+ reductase and cytochrome c was recognized. |
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