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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Cytochromes c / chemistry
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Electrophoresis, Polyacrylamide Gel
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Escherichia coli / genetics
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Ferredoxin-NADP Reductase / chemistry
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Ferredoxins / biosynthesis
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Ferredoxins / chemistry
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Ferredoxins / genetics*
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Ferrous Compounds / chemistry
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Genetic Vectors / genetics
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Organometallic Compounds / chemistry
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Organophosphorus Compounds / chemistry
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Oxidation-Reduction
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Proteomics / methods*
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Pyridines / chemistry
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Quaternary Ammonium Compounds / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification
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Ruthenium / chemistry
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Sequence Analysis, Protein / methods*
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Spectrometry, Mass, Electrospray Ionization
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Spectrophotometry
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Spectrophotometry, Ultraviolet
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Sulfides / chemistry
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Thermus thermophilus / genetics*
Substances
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2-(4-isothiocyanatophenoxy)-1,3,2-dioxaphosphinene 2-oxide
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Ferredoxins
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Ferrous Compounds
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Organometallic Compounds
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Organophosphorus Compounds
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Pyridines
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Quaternary Ammonium Compounds
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Recombinant Proteins
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Sulfides
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apoferredoxin
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Ruthenium
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Cytochromes c
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ammonium ferrous sulfate
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Ferredoxin-NADP Reductase
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2,2',2''-terpyridine
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sodium sulfide