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Overexpression, purification, crystallization and data collection of Sulfolobus solfataricus Sso6206, a novel highly conserved protein.

McEwan AR, Liu H, Oke M, Carter L, Powers H, Dorward M, McMahon SA, White MF, Naismith JH - Acta Crystallogr. Sect. F Struct. Biol. Cryst. Commun. (2006)

Bottom Line: The protein crystallizes in space group P6(1/5)22, with unit-cell parameters a = b = 157.8, c = 307.3 A.Crystals of selenomethionine-variant protein have not yet been obtained.Interestingly, crystal packing, gel filtration and mass spectrometry all suggest the native protein forms a multi-subunit oligomer consisting of >9 subunits.

View Article: PubMed Central - HTML - PubMed

Affiliation: Centre for Biomolecular Sciences, The University, St Andrews KY16 9ST, Scotland.

ABSTRACT
Sso6206, a 10.5 kDa protein from Sulfolobus solfataricus, has been overexpressed, purified and crystallized. The protein crystallizes in space group P6(1/5)22, with unit-cell parameters a = b = 157.8, c = 307.3 A. The crystals are hexagonal bipyramids and a data set has been collected to 2.4 A resolution. Molecular replacement cannot be attempted as no convincing model can be identified. Crystals of selenomethionine-variant protein have not yet been obtained. Interestingly, crystal packing, gel filtration and mass spectrometry all suggest the native protein forms a multi-subunit oligomer consisting of >9 subunits.

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Sequence alignment of Sso6206 and its homologues from archaea and bacteria. The four conserved acidic residues are highlighted in red and the conserved histidine in blue. Sso, Sso6206 from S. solfataricus; Pto, Mka, Pae, Pfu, Mth, Mma, Dps and Sfr, homologues from Picrophilus torridus, Methanopyrus kandleri, Pyrobaculum aerophilum, Pyrococcus furiosus, Methanothermobacter thermautotrophicum, Methanococcus maripaludis, Desulfotalea psychrophila and Shewanella frigidimarina, respectively. Sso6206 contains a single cysteine residue.
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fig1: Sequence alignment of Sso6206 and its homologues from archaea and bacteria. The four conserved acidic residues are highlighted in red and the conserved histidine in blue. Sso, Sso6206 from S. solfataricus; Pto, Mka, Pae, Pfu, Mth, Mma, Dps and Sfr, homologues from Picrophilus torridus, Methanopyrus kandleri, Pyrobaculum aerophilum, Pyrococcus furiosus, Methanothermobacter thermautotrophicum, Methanococcus maripaludis, Desulfotalea psychrophila and Shewanella frigidimarina, respectively. Sso6206 contains a single cysteine residue.

Mentions: Sso6206 is a small acidic protein (MW 10 483 Da, pI 4.5) encoded in the genome of the crenarchaeon Sulfolobus solfataricus (She et al., 2001 ▶) with an unknown function. It is nearly ubiquitous in the archaea, including crenarchaea and euryarchaea, hyperthermophiles, halophiles, psychrophiles and methanogens. In addition, homologues have been annotated in two bacterial species, Shewanella frigidimarina, a marine member of the gamma proteobacteria (Bowman et al., 1997 ▶), and Desulfotalea psychrophila, a sulfate-reducing delta proteobacterium from arctic sediments (Knoblauch et al., 1999 ▶). The limited distribution in bacteria suggests that these two species have obtained the gene by lateral gene transfer from an archaeon. The amino-acid sequence of the protein is highly conserved (Fig. 1 ▶), with greater than 40% identity observed between archaeal family members. Perhaps significantly, there are four conserved acidic residues and a conserved histidine (coloured red and blue, respectively, in Fig. 1 ▶), which are often ligands for bound metal ions. The strong conservation hints at a role for this protein in the archaea, making it an interesting target for structural studies. The protein belongs to Pfam2680, a family which has yet to be characterized. Here, we report the overexpression, crystallization and native data collection of recombinant Sso6206.


Overexpression, purification, crystallization and data collection of Sulfolobus solfataricus Sso6206, a novel highly conserved protein.

McEwan AR, Liu H, Oke M, Carter L, Powers H, Dorward M, McMahon SA, White MF, Naismith JH - Acta Crystallogr. Sect. F Struct. Biol. Cryst. Commun. (2006)

Sequence alignment of Sso6206 and its homologues from archaea and bacteria. The four conserved acidic residues are highlighted in red and the conserved histidine in blue. Sso, Sso6206 from S. solfataricus; Pto, Mka, Pae, Pfu, Mth, Mma, Dps and Sfr, homologues from Picrophilus torridus, Methanopyrus kandleri, Pyrobaculum aerophilum, Pyrococcus furiosus, Methanothermobacter thermautotrophicum, Methanococcus maripaludis, Desulfotalea psychrophila and Shewanella frigidimarina, respectively. Sso6206 contains a single cysteine residue.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2197187&req=5

fig1: Sequence alignment of Sso6206 and its homologues from archaea and bacteria. The four conserved acidic residues are highlighted in red and the conserved histidine in blue. Sso, Sso6206 from S. solfataricus; Pto, Mka, Pae, Pfu, Mth, Mma, Dps and Sfr, homologues from Picrophilus torridus, Methanopyrus kandleri, Pyrobaculum aerophilum, Pyrococcus furiosus, Methanothermobacter thermautotrophicum, Methanococcus maripaludis, Desulfotalea psychrophila and Shewanella frigidimarina, respectively. Sso6206 contains a single cysteine residue.
Mentions: Sso6206 is a small acidic protein (MW 10 483 Da, pI 4.5) encoded in the genome of the crenarchaeon Sulfolobus solfataricus (She et al., 2001 ▶) with an unknown function. It is nearly ubiquitous in the archaea, including crenarchaea and euryarchaea, hyperthermophiles, halophiles, psychrophiles and methanogens. In addition, homologues have been annotated in two bacterial species, Shewanella frigidimarina, a marine member of the gamma proteobacteria (Bowman et al., 1997 ▶), and Desulfotalea psychrophila, a sulfate-reducing delta proteobacterium from arctic sediments (Knoblauch et al., 1999 ▶). The limited distribution in bacteria suggests that these two species have obtained the gene by lateral gene transfer from an archaeon. The amino-acid sequence of the protein is highly conserved (Fig. 1 ▶), with greater than 40% identity observed between archaeal family members. Perhaps significantly, there are four conserved acidic residues and a conserved histidine (coloured red and blue, respectively, in Fig. 1 ▶), which are often ligands for bound metal ions. The strong conservation hints at a role for this protein in the archaea, making it an interesting target for structural studies. The protein belongs to Pfam2680, a family which has yet to be characterized. Here, we report the overexpression, crystallization and native data collection of recombinant Sso6206.

Bottom Line: The protein crystallizes in space group P6(1/5)22, with unit-cell parameters a = b = 157.8, c = 307.3 A.Crystals of selenomethionine-variant protein have not yet been obtained.Interestingly, crystal packing, gel filtration and mass spectrometry all suggest the native protein forms a multi-subunit oligomer consisting of >9 subunits.

View Article: PubMed Central - HTML - PubMed

Affiliation: Centre for Biomolecular Sciences, The University, St Andrews KY16 9ST, Scotland.

ABSTRACT
Sso6206, a 10.5 kDa protein from Sulfolobus solfataricus, has been overexpressed, purified and crystallized. The protein crystallizes in space group P6(1/5)22, with unit-cell parameters a = b = 157.8, c = 307.3 A. The crystals are hexagonal bipyramids and a data set has been collected to 2.4 A resolution. Molecular replacement cannot be attempted as no convincing model can be identified. Crystals of selenomethionine-variant protein have not yet been obtained. Interestingly, crystal packing, gel filtration and mass spectrometry all suggest the native protein forms a multi-subunit oligomer consisting of >9 subunits.

Show MeSH