Biochemical and structural characterization of the hypoxanthine-guanine-xanthine phosphoribosyltransferase from Pyrococcus horikoshii

Dantas, Deyse de Souza; Santos, C.R.; Pereira, Gonçalo Amarante Guimarães; Medrano, Francisco J.

doi:10.1016/j.bbapap.2008.03.006

Cited by 6 publications

(8 citation statements)

References 44 publications

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“…The first Brazilian ANA consensus established a decision tree with the focus on the morphological criteria for pattern recognition and aimed to provide guidance for reading slides ( 8 ). The second Brazilian ANA consensus extended the decision tree and added some considerations for mixed patterns as well as the relevant clinical associations for each pattern ( 9 ). The third ANA consensus included new patterns and some recommendations for quality controls ( 10 ).…”

Section: Discussionmentioning

confidence: 99%

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Report of the First International Consensus on Standardized Nomenclature of Antinuclear Antibody HEp-2 Cell Patterns 2014–2015

et al. 2015

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During the 12th International Workshop on Autoantibodies and Autoimmunity held in Sao Paulo, Brazil, on August 28, 2014, a full day session was devoted to establishing a consensus on the nomenclature of staining patterns observed in the antinuclear antibody (ANA) indirect immunofluorescence test on HEp-2 cells. The current report summarizes the collective agreements with input from the host Brazilian and international communities that represented research, clinical, and diagnostic service laboratories. Patterns are categorized in three major groups (nuclear, cytoplasmic, and mitotic patterns) and each pattern has been defined and described in detail. The consensus nomenclature and representative patterns are made available online at the international consensus on antinuclear antibody pattern (ICAP) website (). To facilitate continuous improvement and input, specific comments on ICAP are encouraged and these will be discussed in subsequent ICAP meetings. The ultimate goal with the establishment of the ICAP is to promote harmonization and understanding of autoantibody test nomenclature, as well as interpretation guidelines for ANA testing, thereby optimizing usage in patient care.

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Section: Discussionmentioning

confidence: 99%

“…Brazil has a long history of ANA pattern consensus with their first workshop held in Goiania in August, 2000. Their first consensus report was published in 2001 ( 8 ) and two subsequent ANA consensus reports published in 2003 ( 9 ) and 2009 ( 10 ). These earlier publications were in Portuguese with some including English abstracts.…”

Section: Introductionmentioning

confidence: 99%

Report of the First International Consensus on Standardized Nomenclature of Antinuclear Antibody HEp-2 Cell Patterns 2014–2015

et al. 2015

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“…However, Tg UPRT has also been described as monomeric or tetrameric (in the presence of GTP, which affects the oligomerization state in solution) . It seems that the oligomerization state of the PRTs could be essential in catalysis and stabilization of the active conformation as well as in thermal stability of the protein …”

Section: Resultsmentioning

confidence: 87%

Enzymatic Production of Non‐Natural Nucleoside‐5′‐Monophosphates by a Thermostable Uracil Phosphoribosyltransferase

et al. 2017

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The use of enzymes as biocatalysts applied to synthesis of modified nucleoside‐5′‐monophosphates (NMPs) is an interesting alternative to traditional multistep chemical methods which offers several advantages, such as stereo‐, regio‐, and enantioselectivity, simple downstream processing, and mild reaction conditions. Herein we report the recombinant expression, production, and purification of uracil phosphoribosyltransferase from Thermus themophilus HB8 (TtUPRT). The structure of TtUPRT has been determined by protein crystallography, and its substrate specificity and biochemical characteristics have been analyzed, providing new structural insights into the substrate‐binding mode. Biochemical characterization of the recombinant protein indicates that the enzyme is a homotetramer, with activity and stability across a broad range of temperatures (50–80 °C), pH (5.5–9) and ionic strength (0–500 mm NaCl). Surprisingly, TtUPRT is able to recognize several 5 and 6‐substituted pyrimidines as substrates. These experimental results suggest TtUPRT could be a valuable biocatalyst for the synthesis of modified NMPs.

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“…As it is expected from thermophilic enzymes, TtXPRT is very active at high temperatures; unfortunately, there is not any described other strict xanthine phosphoribosyltransferase from thermophiles to compare these data. The most similar enzymes are HGXPRTs from thermoacidophile archaeon Sulfolobus solfataricus and hyperthermophilic archaeon Pyrococcus horikoshii, which are active at high temperatures (Hansen et al 2014;de Souza et al 2008). Regarding the effect of pH on enzyme activity, all described specific XPRTs work reasonably better in neutral-alkaline conditions, such as XPRTs from Streptococcus faecalis (Miller et al 1974) and E. coli (Vos et al 1997).…”

Section: Biochemical Characterizationmentioning

confidence: 99%

Cloning, expression and biochemical characterization of xanthine and adenine phosphoribosyltransferases fromThermus thermophilusHB8

Arco

Martínez

Donday

et al. 2017

Biocatalysis and Biotransformation

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Purine phosphoribosyltransferases, purine PRTs, are essential enzymes in the purine salvage pathway of living organisms. They are involved in the formation of C-N glycosidic bonds in purine nucleosides-5 0 -monophosphate (NMPs) through the transfer of the 5-phosphoribosyl group from 5-phospho-a-D-ribosyl-1-pyrophosphate (PRPP) to purine nucleobases in the presence of Mg 2þ . Herein, we report a simple and thermostable process for the one-pot, one-step synthesis of some purine NMPs using xanthine phosphoribosyltransferase, XPRT or adenine phosphoribosyltransferase, APRT2, from Thermus thermophilus HB8. In this sense, the cloning, expression and purification of TtXPRT and TtAPRT2 is described for the first time. Both genes, xprt and aprt2 were expressed as his-tagged enzymes in E. coli BL21(DE3) and purified by a heat-shock treatment, followed by Ni-affinity chromatography and a final, polishing gel-filtration chromatography. Biochemical characterization revealed TtXPRT as a tetramer and TtAPRT2 as a dimer. In addition, both enzymes displayed a strong temperature dependence (relative activity >75% in a temperature range from 70 to 90 C), but they also showed very different behaviour under the influence of pH. While TtXPRT is active in a pH range from 5 to 7, TtAPRT2 has a high dependence of alkaline conditions, showing highest activity values in a pH range from 8 to 10. Finally, substrate specificity studies were performed in order to explore their potential as industrial biocatalyst for NMPs synthesis. ARTICLE HISTORY

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Biochemical and structural characterization of the hypoxanthine-guanine-xanthine phosphoribosyltransferase from Pyrococcus horikoshii

Cited by 6 publications

References 44 publications

Report of the First International Consensus on Standardized Nomenclature of Antinuclear Antibody HEp-2 Cell Patterns 2014–2015

Report of the First International Consensus on Standardized Nomenclature of Antinuclear Antibody HEp-2 Cell Patterns 2014–2015

Enzymatic Production of Non‐Natural Nucleoside‐5′‐Monophosphates by a Thermostable Uracil Phosphoribosyltransferase

Cloning, expression and biochemical characterization of xanthine and adenine phosphoribosyltransferases fromThermus thermophilusHB8

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