Crystal Structure of the Alcohol Dehydrogenase from the Hyperthermophilic Archaeon Sulfolobus solfataricus at 1.85Å Resolution

Esposito, Luciana; Sica, F.; Raia, Carlo A.; Giordano, Antonietta; Rossi, Mosè; Mazzarella, L.; Zagari, Adriana

doi:10.1016/s0022-2836(02)00088-8

Cited by 92 publications

(108 citation statements)

References 49 publications

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“…The results of the present study document that N-⑀-methylation of specific lysyl residues of ␤-glycosidase from S. solfataricus is associated with increased stability toward high temperature and SDS as well as a lower susceptibility to denaturation and aggregation, particularly at the high temperatures (Ͼ85°C) typical of the natural environment of the organism (31). Such findings are particularly interesting, in light of recent observations collected with amyloid peptide ␤- (25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35), which is the shortest synthetic peptide retaining the ␤-sheet fibril-forming capability and toxicity of ␤-amyloid, the peptide believed to cause Alzheimer's disease. N-Methylation of amyloid peptide ␤-(25-35) prevented the aggregation and inhibited the resulting toxicity of the wild type peptide (11).…”

Section: Discussionsupporting

confidence: 69%

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Thermal Stability and Aggregation of Sulfolobus solfataricus β-Glycosidase Are Dependent upon the N-∈-Methylation of Specific Lysyl Residues

Febbraio

Andolfo

Tanfani

et al. 2004

Journal of Biological Chemistry

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Methylation in vivo is a post-translational modification observed in several organisms belonging to eucarya, bacteria, and archaea. Although important implications of this modification have been demonstrated in several eucaryotes, its biological role in hyperthermophilic archaea is far from being understood. The aim of this work is to clarify some effects of methylation on the properties of ␤-glycosidase from Sulfolobus solfataricus, by a structural comparison between the native, methylated protein and its unmethylated counterpart, recombinantly expressed in Escherichia coli. Analysis by Fourier transform infrared spectroscopy indicated similar secondary structure contents for the two forms of the protein. However, the study of temperature perturbation by Fourier transform infrared spectroscopy and turbidimetry evidenced denaturation and aggregation events more pronounced in recombinant than in native ␤-glycosidase. Red Nile fluorescence analysis revealed significant differences of surface hydrophobicity between the two forms of the protein. Unlike the native enzyme, which dissociated into SDS-resistant dimers upon exposure to the detergent, the recombinant enzyme partially dissociated into monomers. By electrospray mapping, the methylation sites of the native protein were identified. A computational analysis of ␤-glycosidase three-dimensional structure and comparisons with other proteins from S. solfataricus revealed analogies in the localization of methylation sites in terms of secondary structural elements and overall topology. These observations suggest a role for the methylation of lysyl residues, located in selected domains, in the thermal stabilization of ␤-glycosidase from S. solfataricus.

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Section: Discussionsupporting

confidence: 69%

“…A recent observation of high medical interest concerns the inhibition of toxicity of the synthetic ␤-amyloid peptide ␤- (25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35) by N-methylation (11).…”

mentioning

confidence: 99%

Thermal Stability and Aggregation of Sulfolobus solfataricus β-Glycosidase Are Dependent upon the N-∈-Methylation of Specific Lysyl Residues

Febbraio

Andolfo

Tanfani

et al. 2004

Journal of Biological Chemistry

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“…In addition, a conserved sequence for a structural zinc binding site DXCXXCXXXXXXXC (where X is any residue) can be found (amino acids 90, 93, 96, and 104). Here, amino acid 90 is changed from cysteine to aspartic acid, as already observed in Sulfolobus solfataricus and Aeropyrum pernix (6,14). Whether the native and active reductases contain one or two zinc ions cannot be decided based on our data.…”

Section: Discussionmentioning

confidence: 59%

3-Hydroxypropionyl-Coenzyme A Dehydratase and Acryloyl-Coenzyme A Reductase, Enzymes of the Autotrophic 3-Hydroxypropionate/4-Hydroxybutyrate Cycle in the Sulfolobales

et al. 2009

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A 3-hydroxypropionate/4-hydroxybutyrate cycle operates in autotrophic CO 2 fixation in various Crenarchaea, as studied in some detail in Metallosphaera sedula. This cycle and the autotrophic 3-hydroxypropionate cycle in Chloroflexus aurantiacus have in common the conversion of acetyl-coenzyme A (CoA) and two bicarbonates via 3-hydroxypropionate to succinyl-CoA. Both cycles require the reductive conversion of 3-hydroxypropionate to propionyl-CoA. In M. sedula the reaction sequence is catalyzed by three enzymes. The first enzyme, 3-hydroxypropionyl-CoA synthetase, catalyzes the CoA-and MgATP-dependent formation of 3-hydroxypropionyl-CoA. The next two enzymes were purified from M. sedula or Sulfolobus tokodaii and studied. 3-Hydroxypropionyl-CoA dehydratase, a member of the enoyl-CoA hydratase family, eliminates water from 3-hydroxypropionyl-CoA to form acryloyl-CoA. Acryloyl-CoA reductase, a member of the zinc-containing alcohol dehydrogenase family, reduces acryloyl-CoA with NADPH to propionyl-CoA. Genes highly similar to the Metallosphaera CoA synthetase, dehydratase, and reductase genes were found in autotrophic members of the Sulfolobales. The encoded enzymes are only distantly related to the respective three enzyme domains of propionyl-CoA synthase from C. aurantiacus, where this trifunctional enzyme catalyzes all three reactions. This indicates that the autotrophic carbon fixation cycles in Chloroflexus and in the Sulfolobales evolved independently and that different genes/enzymes have been recruited in the two lineages that catalyze the same kinds of reactions.

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“…Amino acid composition and its substitution patterns between mesophilic and hyperthermophilic proteins shed light on the understanding of common features of thermostability (56,63). T. guaymasensis ADH showed 77% identity to T. brockii ADH and The information refers to the recent crystal structures (11,19,20,27,30 (35). The uncharged polar residues Gln, Asn, and Ser decreased in T. guaymasensis ADH, in which the first two are prone to deamidation and known to be the most temperature sensitive (14,67).…”

Section: Discussionmentioning

confidence: 99%

“…Zinc-containing ADHs constitute a big protein family with various enzyme activities, including alcohol dehydrogenase, polyol dehydrogenase, and cinnamyl alcohol dehydrogenase activities (37). A large number of zinc-containing ADHs, including those from hyperthermophiles Thermococcus kodakaraensis, Pyrococcus horikoshii, Aeropyrum pernix, and Sulfolobus solfataricus, contain catalytic and structural zincs (5,11,20,25,30). The zinc-containing ADHs from the mesophile Clostridium beijerinckii and thermophiles Thermoanaerobacter brockii and Thermoanaerobacter ethanolicus contain only catalytic zinc (34).…”

mentioning

confidence: 99%

Characterization of a Zinc-Containing Alcohol Dehydrogenase with Stereoselectivity from the Hyperthermophilic Archaeon Thermococcus guaymasensis

Ying

2011

J Bacteriol

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An alcohol dehydrogenase (ADH) from hyperthermophilic archaeon Thermococcus guaymasensis was purified to homogeneity and was found to be a homotetramer with a subunit size of 40 ؎ 1 kDa. The gene encoding the enzyme was cloned and sequenced; this gene had 1,095 bp, corresponding to 365 amino acids, and showed high sequence homology to zinc-containing ADHs and L-threonine dehydrogenases with binding motifs of catalytic zinc and NADP ؉ . Metal analyses revealed that this NADP ؉ -dependent enzyme contained 0.9 ؎ 0.03 g-atoms of zinc per subunit. It was a primary-secondary ADH and exhibited a substrate preference for secondary alcohols and corresponding ketones. Particularly, the enzyme with unusual stereoselectivity catalyzed an anti-Prelog reduction of racemic (R/S)-acetoin to (2R,3R)-2,3-butanediol and meso-2,3-butanediol. The optimal pH values for the oxidation and formation of alcohols were 10.5 and 7.5, respectively. Besides being hyperthermostable, the enzyme activity increased as the temperature was elevated up to 95°C. The enzyme was active in the presence of methanol up to 40% (vol/vol) in the assay mixture. The reduction of ketones underwent high efficiency by coupling with excess isopropanol to regenerate NADPH. The kinetic parameters of the enzyme showed that the apparent K m values and catalytic efficiency for NADPH were 40 times lower and 5 times higher than those for NADP ؉ , respectively. The physiological roles of the enzyme were proposed to be in the formation of alcohols such as ethanol or acetoin concomitant to the NADPH oxidation.

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Crystal Structure of the Alcohol Dehydrogenase from the Hyperthermophilic Archaeon Sulfolobus solfataricus at 1.85Å Resolution

Cited by 92 publications

References 49 publications

Thermal Stability and Aggregation of Sulfolobus solfataricus β-Glycosidase Are Dependent upon the N-∈-Methylation of Specific Lysyl Residues

Thermal Stability and Aggregation of Sulfolobus solfataricus β-Glycosidase Are Dependent upon the N-∈-Methylation of Specific Lysyl Residues

3-Hydroxypropionyl-Coenzyme A Dehydratase and Acryloyl-Coenzyme A Reductase, Enzymes of the Autotrophic 3-Hydroxypropionate/4-Hydroxybutyrate Cycle in the Sulfolobales

Characterization of a Zinc-Containing Alcohol Dehydrogenase with Stereoselectivity from the Hyperthermophilic Archaeon Thermococcus guaymasensis

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