, Volume 224, Issue 3, pp 668–679 | Cite as

Co-occurrence of both l-asparaginase subtypes in Arabidopsis: At3g16150 encodes a K+-dependent l-asparaginase

  • Luanne Bruneau
  • Ralph Chapman
  • Frédéric Marsolais
Original Article


l-asparaginases (EC are hypothesized to play an important role in nitrogen supply to sink tissues, especially in legume-developing seeds. Two plant l-asparaginase subtypes were previously identified according to their K+-dependence for catalytic activity. An l-asparaginase homologous to Lupinus K+-independent enzymes with activity towards β-aspartyl dipeptides, At5g08100, has been previously characterized as a member of the N-terminal nucleophile amidohydrolase superfamily in Arabidopsis. In this study, a K+-dependent l-asparaginase from Arabidopsis, At3g16150, is characterized. The recombinants At3g16150 and At5g08100 share a similar subunit structure and conserved autoproteolytic pentapeptide cleavage site, commencing with the catalytic Thr nucleophile, as determined by ESI-MS. The catalytic activity of At3g16150 was enhanced approximately tenfold in the presence of K+. At3g16150 was strictly specific for l-Asn, and had no activity towards β-aspartyl dipeptides. At3g16150 also had an approximately 80-fold higher catalytic efficiency with l-Asn relative to At5g08100. Among the β-aspartyl dipeptides tested, At5g08100 had a preference for β-aspartyl-His, with catalytic efficiency comparable to that with l-Asn. The phylogenetic analysis revealed that At3g16150 and At5g08100 belong to two distinct subfamilies. The transcript levels of At3g16150 and At5g08100 were highest in sink tissues, especially in flowers and siliques, early in development, as determined by quantitative RT-PCR. The overlapping spatial patterns of expression argue for a partially redundant function of the enzymes. However, the high catalytic efficiency suggests that the K+-dependent enzyme may metabolize l-Asn more efficiently under conditions of high metabolic demand for N.


Arabidopsis l-asparaginase K+-dependent At3g16150 gene Seed nitrogen supply Isoaspartyl dipeptidase 



N-terminal nucleophile




2-(N-morpholino)ethanesulfonic acid


Arabidopsis genome initiative


Arabidopsis Biological Resource Center


Isopropyl β-D-1-thiogalactopyranoside


Day after anthesis


Fresh weight


Diethyl pyrocarbonate


The Arabidopsis Information Resource


National Center for Biotechnology Information


The Institute for Genome Research


Canadian Bioinformatics Resource


Least significant difference


Expressed sequence tag


Human glycosylasparaginase


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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Luanne Bruneau
    • 1
  • Ralph Chapman
    • 1
  • Frédéric Marsolais
    • 1
  1. 1.Agriculture and Agri-Food CanadaSouthern Crop Protection and Food Research CentreLondonCanada

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