Abstract
Proline is a key factor in plant adaptation to environmental stresses. The Δ1-pyrroline-5-carboxylate synthetase catalyzes the first committed step and the rate-limiting step for proline biosynthesis in both plants and mammals. This enzyme catalyzes the reduction of glutamate to pyrroline-5-carboxylate in two sequential steps including the phosphorylation and the reduction of its precursor. Several methods were established to assay P5CS activity but however none of them are fully reliable. Therefore, we developed a new simple and reliable assay which is based on the quantification of Pi. This assay allowed us to determine the optimal pH, the apparent K m and V m of P5CS with regard to ATP and glutamate.
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Acknowledgments
E.P. was supported by a Ph.D. grant from the French “Ministère de l’Education Nationale, France” and M.A.G. by a grant from the French “Ministère des Affaires Etrangères” (French Embassy, Tunis).
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Parre, E. et al. (2010). A New Method for Accurately Measuring Δ1-Pyrroline-5-Carboxylate Synthetase Activity. In: Sunkar, R. (eds) Plant Stress Tolerance. Methods in Molecular Biology, vol 639. Humana Press. https://doi.org/10.1007/978-1-60761-702-0_21
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DOI: https://doi.org/10.1007/978-1-60761-702-0_21
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