Transamination, deamidation, and the utilisation of asparagine amino nitrogen in pea leaves

Abstract

The fate of 15N from the amino group of labelled asparagine was followed in growing pea leaves, in the presence and absence of inhibitors of deamidation (DONV, 5-diazo-4-oxo-L-norvaline) and transamination (AOA, aminooxyacetate). The label was widely incorporated into various amino acids, especially aspartate, glutamate, alanine, and homoserine, as well as glycine and serine. Treatment with AOA considerably decreased the label of all these amino acids except aspartate, consistent with the production of the latter by deamidation of asparagine. This was confirmed by the use of DONV, which decreased aspartate labelling by over 70%; a similar decrease in glutamate labelling suggested that the latter was labelled predominantly by transamination of aspartate. In contrast, DONV had a much smaller effect on the labelling of alanine, homoserine, glycine, and serine, indicating a direct transfer of amino nitrogen from asparagine, rather than transfer from aspartate (or glutamate). The labelling of glycine and serine and the transfer of amino nitrogen to ammonia and glutamine (amide group) were consistent with a flow of asparagine nitrogen into the photorespiratory nitrogen cycle. During leaf expansion there was a decrease in the amount of asparagine metabolised, and a decreasing participation of deamidation as the leaf matured.