PMID:20812989

Janowitz, T, Ajonina, I, Perbandt, M, Woltersdorf, C, Hertel, P, Liebau, E and Gigengack, U (2010) The 3-ureidopropionase of Caenorhabditis elegans, an enzyme involved in pyrimidine degradation.FEBS J.

Abstract

Pyrimidines are important metabolites in all cells. Levels of cellular pyrimidines are controlled by multiple mechanisms, with one of these comprising the reductive degradation pathway. In the model invertebrate Caenorhabditis elegans, two of the three enzymes of reductive pyrimidine degradation have previously been characterized. The enzyme catalysing the final step of pyrimidine breakdown, 3-ureidopropionase (beta-alanine synthase), had only been identified based on homology. We therefore cloned and functionally expressed the 3-ureidopropionase of C. elegans as hexahistidine fusion protein. The purified recombinant enzyme readily converted the two pyrimidine degradation products: 3-ureidopropionate and 2-methyl-3-ureidopropionate. The enzyme showed a broad pH optimum between pH 7.0 and 8.0. Activity was highest at approximately 40 degrees C, although the half-life of activity was only 65 s at that temperature. The enzyme showed clear Michaelis-Menten kinetics, with a K(m) of 147 +/- 26 mum and a V(max) of 1.1 +/- 0.1 U.mg protein(-1). The quaternary structure of the recombinant enzyme was shown to correspond to a dodecamer by 'blue native' gel electrophoresis and gel filtration. The organ specific and subcellular localization of the enzyme was determined using a translational fusion to green fluorescent protein and high expression was observed in striated muscle cells. With the characterization of the 3-ureidopropionase, the reductive pyrimidine degradation pathway in C. elegans has been functionally characterized. Structured digital abstract * MINT-7986015: 3-ureidopropionase (uniprotkb:Q19437) and 3-ureidopropionase (uniprotkb:Q19437) bind (MI:0407) by blue native page (MI:0276).

Links

PubMed Online version:10.1111/j.1742-4658.2010.07805.x

Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status

References

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