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Purification, primary structure, and properties of Euphorbia characias latex purple acid phosphatase

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Abstract

A purple acid phosphatase was purified to homogeneity from Euphorbia characias latex. The native protein has a molecular mass of 130 ± 10 kDa and is formed by two apparently identical subunits, each containing one Fe(III) and one Zn(II) ion. The two subunits are connected by a disulfide bridge. The enzyme has an absorbance maximum at 540 nm, conferring a characteristic purple color due to a charge-transfer transition caused by a tyrosine residue (Tyr172) coordinated to the ferric ion. The cDNA nucleotide sequence contains an open reading frame of 1392 bp, and the deduced sequence of 463 amino acids shares a very high degree of identity (92–99%) to other purple acid phosphatases isolated from several higher plants. The enzyme hydrolyzes well p-nitrophenyl phosphate, a typical artificial substrate, and a broad range of natural phosphorylated substrates, such as ATP, ADP, glucose-6-phosphate, and phosphoenolpyruvate. The enzyme displays a pH optimum of 5.75 and is inhibited by molybdate, vanadate, and Zn2+, which are typical acid phosphatase inhibitors.

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Abbreviations

AP:

acid phosphatases

ELPAP:

Euphorbia latex purple acid phosphatase (protein)

PAPs:

purple acid phosphatases

p-NPP:

4-nitrophenyl phosphate

RT-PCR:

reverse transcription-polymerase chain reaction

SDS-PAGE:

sodium dodecyl sulfate polyacrylamide gel electrophoresis

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Authors and Affiliations

  1. Department of Applied Sciences in Biosystems, University of Cagliari, Cagliari, Italy

    F. Pintus, D. Spano, S. Corongiu, G. Floris & R. Medda

Authors
  1. F. Pintus
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  2. D. Spano
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  3. S. Corongiu
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  4. G. Floris
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  5. R. Medda
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Corresponding author

Correspondence to R. Medda.

Additional information

Published in Russian in Biokhimiya, 2011, Vol. 76, No. 6, pp. 849–857.

These authors contributed equally to this work.

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Pintus, F., Spano, D., Corongiu, S. et al. Purification, primary structure, and properties of Euphorbia characias latex purple acid phosphatase. Biochemistry Moscow 76, 694–701 (2011). https://doi.org/10.1134/S0006297911060101

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  • DOI: https://doi.org/10.1134/S0006297911060101

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