(R)-Mevalonate 3-Phosphate Is an Intermediate of the Mevalonate Pathway in Thermoplasma acidophilum

Azami, Yasuhiro; Hattori, Akira; Nishimura, Hiroto; Kawaide, Hiroshi; Yoshimura, Teizo; Hemmi, Hisashi

doi:10.1074/jbc.m114.562686

Cited by 45 publications

(54 citation statements)

References 22 publications

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“…In the modified pathways, isopentenyl diphosphate is formed from isopentenyl phosphate, which is not an intermediate of the classical pathway. However, the biosynthetic routes to convert MVA into isopentenyl phosphate are different among archaeal lineages; for example, MVA 5-kinase and phosphomevalonate decarboxylase are used in a halophilic archaeon Haloferax volcanii (53), and MVA 3-kinase, MVA-3-phosphate 5-kinase, and probably MVA-3,5-bisphosphate decarboxylase are used in a thermophilic archaeon, Thermoplasma acidophilum (54,55). Among these recently discovered enzymes, phosphomevalonate decarboxylase and MVA 3-kinase are closely related to DMD.…”

Section: Discussionmentioning

confidence: 99%

In Vivo Formation of the Protein Disulfide Bond That Enhances the Thermostability of Diphosphomevalonate Decarboxylase, an Intracellular Enzyme from the Hyperthermophilic Archaeon Sulfolobus solfataricus

et al. 2015

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In the present study, the crystal structure of recombinant diphosphomevalonate decarboxylase from the hyperthermophilic archaeon Sulfolobus solfataricus was solved as the first example of an archaeal and thermophile-derived diphosphomevalonate decarboxylase. The enzyme forms a homodimer, as expected for most eukaryotic and bacterial orthologs. Interestingly, the subunits of the homodimer are connected via an intersubunit disulfide bond, which presumably formed during the purification process of the recombinant enzyme expressed in Escherichia coli. When mutagenesis replaced the disulfide-forming cysteine residue with serine, however, the thermostability of the enzyme was significantly lowered. In the presence of ␤-mercaptoethanol at a concentration where the disulfide bond was completely reduced, the wild-type enzyme was less stable to heat. Moreover, Western blot analysis combined with nonreducing SDS-PAGE of the whole cells of S. solfataricus proved that the disulfide bond was predominantly formed in the cells. These results suggest that the disulfide bond is required for the cytosolic enzyme to acquire further thermostability and to exert activity at the growth temperature of S. solfataricus. IMPORTANCEThis study is the first report to describe the crystal structures of archaeal diphosphomevalonate decarboxylase, an enzyme involved in the classical mevalonate pathway. A stability-conferring intersubunit disulfide bond is a remarkable feature that is not found in eukaryotic and bacterial orthologs. The evidence that the disulfide bond also is formed in S. solfataricus cells suggests its physiological importance.

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Section: Discussionmentioning

confidence: 99%

In Vivo Formation of the Protein Disulfide Bond That Enhances the Thermostability of Diphosphomevalonate Decarboxylase, an Intracellular Enzyme from the Hyperthermophilic Archaeon Sulfolobus solfataricus

et al. 2015

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“…Recently, evidence for the existence of yet another pathway for IPP biosynthesis in Archaea has been presented. In T. acidophilum , mevalonate-3-kinase converts mevalonate into mevalonate-3-phosphate, which is then converted into mevalonate-3,5-bisphosphate by mevalonate-3-phosphate-5-kinase [79,80]. Mevolonate-3-kinase was subsequently described in Picrophilus torridus (optimal growth at 60°C and pH 1.8–2 [81]), with sequence analysis suggesting the enzyme to also exist in other members of the Thermoplasmatales , the order that includes these two species [82].…”

Section: Phosphodolichol Biosynthesis In Archaeamentioning

confidence: 99%

Lipid sugar carriers at the extremes: The phosphodolichols Archaea use in N-glycosylation

Eichler

Guan

2017

Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biolog

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N-glycosylation, a post-translational modification whereby glycans are covalently linked to select Asn residues of target proteins, occurs in all three domains of life. Across evolution, the N-linked glycans are initially assembled on phosphorylated cytoplasmically-oriented polyisoprenoids, with polyprenol (mainly C55 undecaprenol) fulfilling this role in Bacteria and dolichol assuming this function in Eukarya and Archaea. The eukaryal and archaeal versions of dolichol can, however, be distinguished on the basis of their length, degree of saturation and by other traits. As is true for many facets of their biology, Archaea, best known in their capacity as extremophiles, present unique approaches for synthesizing phosphodolichols. At the same time, general insight into the assembly and processing of glycan-bearing phosphodolichols has come from studies of the archaeal enzymes responsible. In this review, these and other aspects of archaeal phosphodolichol biology are addressed.

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“…To initially fractionate the lysate, we separated the T. acidophilum lysate by anion exchange chromatography using a HiTrap Q HP column. Ten fractions were collected and assayed for MBD activity18. As shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Initial separation of the T. acidophilum lysate was adapted from the procedure of another research group18. In brief, 10 mL of T. acidophilum lysate was applied to a 1 mL HiTrap Q HP anion exchange column (strong quaternary ammonium anion exchanger) pre-equilibrated with 50 mM sodium phosphate buffer [pH 6.5], connected to an AKTA FPLC system.…”

Section: Methodsmentioning

confidence: 99%

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An Adaptation To Life In Acid Through A Novel Mevalonate Pathway

Vinokur

Cummins

Korman

et al. 2016

Sci Rep

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Extreme acidophiles are capable of growth at pH values near zero. Sustaining life in acidic environments requires extensive adaptations of membranes, proton pumps, and DNA repair mechanisms. Here we describe an adaptation of a core biochemical pathway, the mevalonate pathway, in extreme acidophiles. Two previously known mevalonate pathways involve ATP dependent decarboxylation of either mevalonate 5-phosphate or mevalonate 5-pyrophosphate, in which a single enzyme carries out two essential steps: (1) phosphorylation of the mevalonate moiety at the 3-OH position and (2) subsequent decarboxylation. We now demonstrate that in extreme acidophiles, decarboxylation is carried out by two separate steps: previously identified enzymes generate mevalonate 3,5-bisphosphate and a new decarboxylase we describe here, mevalonate 3,5-bisphosphate decarboxylase, produces isopentenyl phosphate. Why use two enzymes in acidophiles when one enzyme provides both functionalities in all other organisms examined to date? We find that at low pH, the dual function enzyme, mevalonate 5-phosphate decarboxylase is unable to carry out the first phosphorylation step, yet retains its ability to perform decarboxylation. We therefore propose that extreme acidophiles had to replace the dual-purpose enzyme with two specialized enzymes to efficiently produce isoprenoids in extremely acidic environments.

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(R)-Mevalonate 3-Phosphate Is an Intermediate of the Mevalonate Pathway in Thermoplasma acidophilum

Cited by 45 publications

References 22 publications

In Vivo Formation of the Protein Disulfide Bond That Enhances the Thermostability of Diphosphomevalonate Decarboxylase, an Intracellular Enzyme from the Hyperthermophilic Archaeon Sulfolobus solfataricus

In Vivo Formation of the Protein Disulfide Bond That Enhances the Thermostability of Diphosphomevalonate Decarboxylase, an Intracellular Enzyme from the Hyperthermophilic Archaeon Sulfolobus solfataricus

Lipid sugar carriers at the extremes: The phosphodolichols Archaea use in N-glycosylation

An Adaptation To Life In Acid Through A Novel Mevalonate Pathway

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