A New Method for Accurately Measuring Δ1-Pyrroline-5-Carboxylate Synthetase Activity

Autor: Jacques Davy de Virville, Mohamed Ali Ghars, Alain Zachowski, Françoise Cochet, Anne-Sophie Leprince, Luc Richard, Marianne Bordenave, Arnould Savouré, Elodie Parre, Delphine Lefebvre-De Vos
Rok vydání: 2010
Předmět:
Zdroj: Methods in Molecular Biology ISBN: 9781607617013
DOI: 10.1007/978-1-60761-702-0_21
Popis: Proline is a key factor in plant adaptation to environmental stresses. The Delta(1)-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.
Databáze: OpenAIRE