- Rapid and sensitive amino-acid sequencing of cloning Thermus thermophilus HB8 ferredoxin by proteomics.
Rapid and sensitive amino-acid sequencing of cloning Thermus thermophilus HB8 ferredoxin by proteomics.
Journal of proteome research (2004-10-12)
Maki Kaneko, Ryoji Masui, Kojiro Ake, Yukihide Kousumi, Seiki Kuramitsu, Minoru Yamaguchi, Hiroki Kuyama, Eiji Ando, Shigemi Norioka, Takashi Nakazawa, Taka-Aki Okamura, Hitoshi Yamamoto, Norikazu Ueyama
PMID15473686
ABSTRACT
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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