Phylogenetic Analysis and Effect of Heat on Conformational Change of Ferredoxin from Hyperthermophilic Archaeon Pyrococcus sp.KOD1

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概要

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• The gene encoding ferredoxin was cloned from hyperthermophilic archaeon Pyrococcus sp.KOD1, and the nucleotide sequence was determined. The ferredoxin gene encodes a polypeptide of 62 amino acids that incorporates a single[4Fe-4S]cluster with a motif(CXXDXXCXXXCP)in the N-terminal region, where one cysteine residue is replaced by an aspartate residue. Sequence comparison of KOD1 ferredoxin indicated high sequence homology to the ferredoxins from other hyperthermophilic archaea Pyrococcus furiosus(75%), Thermococcus litoralis(67%)and hyperthermophilic bacterium Thermotoga maritima(58%). Based on the sequence information, the phylogenetic tree for the ferredoxins containing single[4Fe-4S]cluster was constructed, which indicates a close evolutionary relationship among hyperthermophiles. Natural ferredoxin was purified from KOD1 cells to homogeneity, and its structure was compared with that of recombinant KOD1 ferredoxin expressed in E.coli.Amino terminal sequences for both natural and recombinant ferredoxins were determined, showing that both ferredoxins lacked the initial methionine. Circular dichroism(CD)spectra of these proteins were distinctly different from each other. Upon heat treatment(90℃, 3h), the structure of recombinant ferredoxin was mostly converted to that of natural ferredoxin, suggesting that exposure to high temperature environment plyas an important role for proper folding of natural KOD1 ferredoxin.

• 社団法人日本生物工学会の論文
• 1998-03-25

著者

• Takagi Masahiro

  School Of Materials Science Japan Advanced Institute Of Science And Technology

• Takagi M

  School Of Materials Science Japan Advanced Institute Of Science And Technology

• FUJIWARA SHINSUKE

  Department of Bioscience, School of Science and Technology, Kwansei Gakuin University

• IMANAKA Tadayuki

  Department of Synthetic and Biological Chemistry, Graduate School of Engineering, Kyoto University

• TAKAGI MASAHIRO

  Department of Biotechnology, Graduate School of Engineering, Osaka University

• Fujiwara S

  School Of Science And Engineering Department Of Life Science Kwansei-gakuin University

• Fujiwara Shinsuke

  Department Of Biotechnology Graduate School Of Engineering Osaka University

• Fujiwara Shinsuke

  Department Of Bioscience School Of Science And Technology Kwansei Gakuin University

• SIDDIQUI MASOOD

  Department of Biotechnology, Graduate School of Engineering, Osaka University

• Takagi M

  Nagasaki Univ. Nagasaki Jpn

• Imanaka Tadayuki

  Department Of Applied Biotechnology Faculty Of Engineering Osaka University

• Fujiwara Shinsuke

  School Of Science And Engineering Department Of Life Science Kwansei-gakuin University

• Siddiqui Masood

  Department Of Biotechnology Graduate School Of Engineering Osaka University

• Takagi Masahiro

  Department Of Biotechnology Faculty Of Engineering Osaka University

• Fujiwara Shinsuke

  Department Of Bioscience Graduate School Of Science And Technology Kwansei-gakuin University:researc

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