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Characterisation of the components of the thioredoxin system in the archaeon Sulfolobus solfataricus

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Abstract

The thioredoxin system is a redox machinery widely distributed in nature and involved in several cellular functions. It is constituted of thioredoxin reductase (Trx-B), its protein substrate thioredoxin (Trx-A) and NADPH. We have previously characterised a Trx-B from the hyperthermophile Sulfolobus solfataricus (SsTrx-B3) (Ruocco et al. in Biochimie 86:883–892, 2004). As in the genome of this archaeon, the gene coding for another Trx-B (SsTrx-B2) and for two Trx-A (SsTrx-A1, SsTrx-A2) have been putatively identified, these proteins were obtained as recombinant forms and characterised. SsTrx-B2, different from SsTrx-B3, did not elicit a thioredoxin reductase activity. S. solfataricus possessed only one Trx-B (SsTrx-B3), which had two thioredoxins (SsTrx-A1 and SsTrx-A2) as substrates. These latter showed a homodimeric structure and catalysed insulin reduction using either DTT or NADPH/SsTrx-B3 as electron donors. In addition, the electron transfer between SsTrx-B3 and either SsTrx-A1 or SsTrx-A2 was fully reversible, thus allowing the determination of the redox potential of the thioredoxin system in S. solfataricus. Among the two thioredoxins, SsTrx-A2 appeared slightly more active and stable than SsTrx-A1. These data, besides shedding light on thioredoxin system in S. solfataricus, will contribute to add further information on this key enzyme system in Archaea.

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Abbreviations

Trx-A:

Thioredoxin

Trx-B:

Thioredoxin reductase

Ss :

Sulfolobus solfataricus

DTNB:

5,5'-Dithio-bis(2-nitrobenzoic acid)

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Authors and Affiliations

  1. Dipartimento di Scienze Farmacobiologiche, Università di Catanzaro “Magna Graecia”, “Complesso Ninì Barbieri”, 88021, Roccelletta di Borgia, Catanzaro, Italy

    Pasquale Grimaldi, Immacolata Ruggiero & Mariorosario Masullo

  2. Dipartimento di Biochimica e Biotecnologie Mediche, Università di Napoli Federico II, Via S. Pansini 5, 80131, Napoli, Italy

    Pasquale Grimaldi, Maria Rosaria Ruocco, Immacolata Ruggiero, Paolo Arcari & Mariorosario Masullo

  3. Istituto di Biostrutture e Bioimmagini, CNR, Via Mezzocannone 16, 80134, Napoli, Italy

    Maria Angela Lanzotti, Alessia Ruggiero & Luigi Vitagliano

  4. Dipartimento delle Scienze Biologiche, Sezione di Biostrutture, Università di Napoli Federico II, Via Mezzocannone 16, 80134, Napoli, Italy

    Maria Angela Lanzotti

  5. CEINGE Biotecnologie Avanzate s.c.a r.l, Via Comunale Margherita 482, 80145, Napoli, Italy

    Paolo Arcari

Authors
  1. Pasquale Grimaldi
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  2. Maria Rosaria Ruocco
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  3. Maria Angela Lanzotti
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  4. Alessia Ruggiero
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  5. Immacolata Ruggiero
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  6. Paolo Arcari
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  7. Luigi Vitagliano
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  8. Mariorosario Masullo
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Corresponding author

Correspondence to Mariorosario Masullo.

Additional information

Communicated by G. Antranikian.

Pasquale Grimaldi and Maria Rosaria Ruocco contributed equally to this work.

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Grimaldi, P., Ruocco, M.R., Lanzotti, M.A. et al. Characterisation of the components of the thioredoxin system in the archaeon Sulfolobus solfataricus . Extremophiles 12, 553–562 (2008). https://doi.org/10.1007/s00792-008-0161-y

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  • DOI: https://doi.org/10.1007/s00792-008-0161-y

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