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Abstract
Glutamine Synthetase of Hevea Brasilienis Latex: Characterisation
and Physiological Regulation
Regular exploitation of rubber tree causes intense
metabolic activity in latex cells located in the phloem, associated with
the regeneration of the latex cellular components removed by tapping. In
this context, glutamine synthetase (GS) plays a prominent role by ensuring
ammonium fixation necessary for the regeneration of latex nitrogenous components.
In latex, GS activity was detected in the cytosol only. The enzyme was purified
and its main physico-chemical characteristics were determined. Its molecular
mass was 450 kDa for the native form and 45 kDa ± 5 kDa for the sub-unit. The pH optimum was found to be 8.0 and variations
in the physiological pH range (6.4 to 7.2) strongly affected GS activity.
Km for ammonium, glutamate and ATP were estimated at 8 µM
± 2 µM, 4 mM ± 0.5
mM and 0.6 mM ± 0.05 mM, respectively. Comparison
with the cytosol physiological parameters showed the importance of ATP concentration
for GS activity regulation. Functioning of the purified enzyme in a paraphysiological
(ultrafiltrated, protein-free) medium, as compared with an optimised control
medium, demonstrated that physiological ATP concentrations were significantly
limiting for GS potential activity.
Treatment with ethephon increases latex production
both by increasing latex flow duration and by stimulating the regeneration of
metabolism. In this paper we demonstrated that ethephon treatment induced a
concomitant increase in pH, ATP concentration and GS activity. These results
suggest that biochemical mechanisms involving pH and ATP participate in the
regulation of GS activity in response to ethylene, in addition to the
previously observed stimulation of gene expression.

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