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APRT from erythrocytes of HGPRT deficient patients: Kinetic, regulatory and thermostability properties

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Abstract

Adenine phosphoribosyltransferase (APRT) has been 1200-fold purified from erythrocytes of a patient with partial hipoxanthine-guanine phosphoribosyltransferase (HGPRT) deficiency, Propositus, and in those of a controlHPRT+, with 20% efficiency in both proteins and specific activity of 550 and 243 nmol/h/mgprotein. The specific activity determined in the Propositus enzyme was, in all purification steps, higher than that of the controlHPRT+. Significant changes were found in their thermal stabilities. Half inactivation times at each temperature studied are greater for the Propositus enzyme in the temperature interval 60–80°C. No significant difference has been observed in the affinity constants for adenine and PRPP substrates. Studies on inhibition by the reaction product suggest that AMP is a competitive inhibitor with respect to PRPP in both enzymes, with Ki values of 150 μM in Propositus and 220 μM in controlHPRT+.

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Authors and Affiliations

  1. Instituto de Investigaciones Biomédicas del C.S.I.C., Madrid, Spain

    Javier Crespillo, Pilar Llorente & Luisa Argom´niz

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  1. Javier Crespillo
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  2. Pilar Llorente
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  3. Luisa Argom´niz
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Crespillo, J., Llorente, P. & Argom´niz, L. APRT from erythrocytes of HGPRT deficient patients: Kinetic, regulatory and thermostability properties. Mol Cell Biochem 254, 359–363 (2003). https://doi.org/10.1023/A:1027323521969

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  • DOI: https://doi.org/10.1023/A:1027323521969

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