Functional properties of the protein disulfide oxidoreductase from the archaeon Pyrococcus furiosus: a member of a novel protein family related to protein disulfide-isomerase.

Protein disulfide oxidoreductases are ubiquitous redox enzymes that catalyse dithiol-disulfide exchange reactions with a CXXC sequence motif at their active site. A disulfide oxidoreductase, a highly thermostable protein, was isolated from Pyrococcus furiosus (PfPDO), which is characterized by two redox sites (CXXC) and an unusual molecular mass. Its 3D structure at high resolution suggests that it may be related to the multidomain protein disulfide-isomerase (PDI), which is currently known only in eukaryotes. This work focuses on the functional characterization of PfPDO as well as its relation to the eukaryotic PDIs. Assays of oxidative, reductive, and isomerase activities of PfPDO were performed, which revealed that the archaeal protein not only has oxidative and reductive activity, but also isomerase activity. On the basis of structural data, two single mutants (C35S and C146S) and a double mutant (C35S/C146S) of PfPDO were constructed and analyzed to elucidate the specific roles of the two redox sites. The results indicate that the CPYC site in the C-terminal half of the protein is fundamental to reductive/oxidative activity, whereas isomerase activity requires both active sites. In comparison with PDI, the ATPase activity was tested for PfPDO, which was found to be cation-dependent with a basic pH optimum and an optimum temperature of 90 degrees C. These results and an investigation on genomic sequence databases indicate that PfPDO may be an ancestor of the eukaryotic PDI and belongs to a novel protein disulfide oxidoreductase family.

[1]  K. Ishikawa,et al.  A hyperthermostable novel protein-disulfide oxidoreductase is reduced by thioredoxin reductase from hyperthermophilic archaeon Pyrococcus horikoshii. , 2003, Archives of biochemistry and biophysics.

[2]  E. Koonin,et al.  Comparative genomics of archaea: how much have we learned in six years, and what's next? , 2003, Genome Biology.

[3]  David Eisenberg,et al.  Genomic evidence that the intracellular proteins of archaeal microbes contain disulfide bonds , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[4]  Jian Wang,et al.  A complete sequence of the T. tengcongensis genome. , 2002, Genome research.

[5]  D. Wishart,et al.  Identification of a novel archaebacterial thioredoxin: determination of function through structure. , 2002, Biochemistry.

[6]  Peter Klappa,et al.  Protein disulfide isomerases exploit synergy between catalytic and specific binding domains , 2002, EMBO reports.

[7]  J. Johnson,et al.  Characterization of the nucleotide-binding capacity and the ATPase activity of the PIP3-binding protein JFC1 , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[8]  Mark A. Ragan,et al.  The complete genome of the crenarchaeon Sulfolobus solfataricus P2 , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[9]  D E Wemmer,et al.  Solution nuclear magnetic resonance structure of a protein disulfide oxidoreductase from Methanococcus jannaschii , 2001, Protein science : a publication of the Protein Society.

[10]  I Uchiyama,et al.  Comparison between Pyrococcus horikoshii and Pyrococcus abyssi genome sequences reveals linkage of restriction-modification genes with large genome polymorphisms. , 2000, Gene.

[11]  K Watanabe,et al.  Archaeal adaptation to higher temperatures revealed by genomic sequence of Thermoplasma volcanium. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[12]  L. Cerchia,et al.  The Chromosomal Protein Sso7d of the CrenarchaeonSulfolobus solfataricus Rescues Aggregated Proteins in an ATP Hydrolysis-dependent Manner* , 2000, The Journal of Biological Chemistry.

[13]  Dmitrij Frishman,et al.  The genome sequence of the thermoacidophilic scavenger Thermoplasma acidophilum , 2000, Nature.

[14]  K. Kim,et al.  A thioredoxin from the hyperthermophilic archaeon Methanococcus jannaschii has a glutaredoxin-like fold but thioredoxin-like activities. , 2000, Biochemistry.

[15]  R. Freedman.  Novel disulfide oxidoreductase in search of a function , 1998, Nature Structural Biology.

[16]  R. Ladenstein,et al.  A protein disulfide oxidoreductase from the archaeon Pyrococcus furiosus contains two thioredoxin fold units , 1998, Nature Structural Biology.

[17]  R. Huber,et al.  The complete genome of the hyperthermophilic bacterium Aquifex aeolicus , 1998, Nature.

[18]  L. Pearl,et al.  Identification and Structural Characterization of the ATP/ADP-Binding Site in the Hsp90 Molecular Chaperone , 1997, Cell.

[19]  R. Ladenstein,et al.  Crystallization and Preliminary X-Ray Structure Analysis of a Hyperthermostable Thioltransferase from the ArchaeonPyrococcus furiosus , 1997 .

[20]  A. Nicolas,et al.  An atypical topoisomerase II from archaea with implications for meiotic recombination , 1997, Nature.

[21]  T. Creighton,et al.  Identifying and characterizing a structural domain of protein disulfide isomerase. , 1996, Biochemistry.

[22]  M Nilges,et al.  Structure determination of the N-terminal thioredoxin-like domain of protein disulfide isomerase using multidimensional heteronuclear 13C/15N NMR spectroscopy. , 1996, Biochemistry.

[23]  L. Ruddock,et al.  pH-dependence of the dithiol-oxidizing activity of DsbA (a periplasmic protein thiol:disulphide oxidoreductase) and protein disulphide-isomerase: studies with a novel simple peptide substrate. , 1996, The Biochemical journal.

[24]  J. Reinstein,et al.  Assessment of the ATP Binding Properties of Hsp90 (*) , 1996, The Journal of Biological Chemistry.

[25]  P. Gueguen,et al.  ATP Binding and Hydrolysis by the Multifunctional Protein Disulfide Isomerase (*) , 1996, The Journal of Biological Chemistry.

[26]  M. Rossi,et al.  The Purification, Cloning, and High Level Expression of a Glutaredoxin-like Protein from the Hyperthermophilic Archaeon Pyrococcus furiosus (*) , 1995, The Journal of Biological Chemistry.

[27]  L. Cerchia,et al.  The chaperonin from the archaeon Sulfolobus solfataricus promotes correct refolding and prevents thermal denaturation in vitro , 1994, Protein science : a publication of the Protein Society.

[28]  M. Rossi,et al.  A thioredoxin from the extreme thermophilic archaeon Sulfolobus solfataricus , 1994 .

[29]  E. Quemeneur,et al.  A major phosphoprotein of the endoplasmic reticulum is protein disulfide isomerase. , 1994, The Journal of biological chemistry.

[30]  L. Vuillard,et al.  Silver-staining of proteins in polyacrylamide gels: a general overview. , 1994, Cellular and molecular biology.

[31]  T. Creighton,et al.  Folding in vitro of bovine pancreatic trypsin inhibitor in the presence of proteins of the endoplasmic reticulum , 1992, Proteins.

[32]  H. Hogenkamp,et al.  The purification, characterization, and primary structure of a small redox protein from Methanobacterium thermoautotrophicum, an archaebacterium. , 1992, The Journal of biological chemistry.

[33]  K. Vuori,et al.  Expression and site-directed mutagenesis of human protein disulfide isomerase in Escherichia coli. This multifunctional polypeptide has two independently acting catalytic sites for the isomerase activity. , 1992, The Journal of biological chemistry.

[34]  P. R. Sibbald,et al.  The P-loop--a common motif in ATP- and GTP-binding proteins. , 1990, Trends in biochemical sciences.

[35]  R. Freedman,et al.  Kinetics and specificity of homogeneous protein disulphide-isomerase in protein disulphide isomerization and in thiol-protein-disulphide oxidoreduction. , 1983, The Biochemical journal.

[36]  D. Barker.  Cloning and amplified expression of the tyrosyl-tRNA synthetase genes of Bacillus stearothermophilus and Escherichia coli. , 1982, European journal of biochemistry.

[37]  T. Creighton,et al.  Counting integral numbers of amino groups per polypeptide chain , 1980, FEBS letters.

[38]  P. A. Lanzetta,et al.  An improved assay for nanomole amounts of inorganic phosphate. , 1979, Analytical biochemistry.

[39]  A. Holmgren,et al.  Thioredoxin catalyzes the reduction of insulin disulfides by dithiothreitol and dihydrolipoamide. , 1979, The Journal of biological chemistry.

[40]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[41]  G R Stark,et al.  Use of dimethyl suberimidate, a cross-linking reagent, in studying the subunit structure of oligomeric proteins. , 1970, Proceedings of the National Academy of Sciences of the United States of America.

[42]  M. Kunitz.  A spectrophotometric method for the measurement of ribonuclease activity. , 1946, The Journal of biological chemistry.

[43]  M. Rossi,et al.  Protein disulfide oxidoreductase from Pyrococcus furiosus: biochemical properties. , 2001, Methods in enzymology.

[44]  C. Fraser,et al.  Genome of Thermotoga maritima MSB8. , 2001, Methods in enzymology.

[45]  R. Ladenstein,et al.  Protein disulfide oxidoreductase from Pyrococcus furiosus: structural properties. , 2001, Methods in enzymology.

[46]  Y. Kawarabayasi,et al.  Complete genome sequence of an aerobic thermoacidophilic crenarchaeon, Sulfolobus tokodaii strain7. , 2001, DNA research : an international journal for rapid publication of reports on genes and genomes.

[47]  James R. Brown,et al.  Genomic sequence of hyperthermophile, Pyrococcus furiosus: implications for physiology and enzymology. , 2001, Methods in enzymology.

[48]  Toshihiro Tanaka,et al.  Complete Genome Sequence of an Aerobic Hyper-thermophilic Crenarchaeon, Aeropyrum pernix K1 (Supplement) , 1999 .

[49]  Y. Kawarabayasi,et al.  Complete genome sequence of an aerobic hyper-thermophilic crenarchaeon, Aeropyrum pernix K1. , 1999, DNA research : an international journal for rapid publication of reports on genes and genomes.

[50]  J. Baross,et al.  Overview of hyperthermophiles and their heat-shock proteins. , 1996, Advances in protein chemistry.

[51]  R. Saiki.  2 – AMPLIFICATION OF GENOMIC DNA , 1990 .

[52]  Thomas A. Kunkel,et al.  Rapid and efficient site-specific mutagenesis without phenotypic selection. , 1985, Proceedings of the National Academy of Sciences of the United States of America.