Purification and characterization of phosphoribosylpyrophosphate synthetase from rubber tree latex
Phosphoribosylpyrophosphate synthetase (PRS; Ec 2.7.6.1) from #Hevea brasiliensis# Müll. Arg. latex was located in the cytosol. After purification, its apparent molecular weight under nondenaturing conditions was estimated at 200,000 + or - 10,000; a single band at 57,000 + or - 3,000 was detected after sodium dodecyl sulfate. Polyacrylamide gel electrophoresis. The enzyme seemed to be a homotetramer. Its affinity constants were estimated at 200 + or - 30 PM Dr adenosine triphosphate and 40 + or - 2 PM for ribose.5.phosphate . The purified enzyme proved to be functional in a paraphysiological medium (cytosol deproteinized by ultrafiltration). Optimum pH was 7.5 in buffer and 6.5 in a paraphysiological medium. No PRS activity vas detected in the absence of the Mg2+ ion. Of the numerous compounds tested, only Mn2+, phosphoribosylpyrophosphate, and inorganic phosphate affected the enzymatic reaction. Mn2+ (inhibitor constant = 20 PM) and phosphoribosylpyrophosphate (inhibitor constant = 30 PM) were inhibitors. PRS responded allosterically (Hill's coefficient = 2.3) to ribulose-5-phosphate in the presence of physiological concentration of inorganic phosphate (10 mM). These results are set in the physiological context of laticifers.
Main Authors: | , , , |
---|---|
Format: | article biblioteca |
Language: | eng |
Subjects: | F62 - Physiologie végétale - Croissance et développement, Hevea brasiliensis, latex, enzyme, pyrophosphate, purification, technique analytique, http://aims.fao.org/aos/agrovoc/c_3589, http://aims.fao.org/aos/agrovoc/c_4214, http://aims.fao.org/aos/agrovoc/c_2603, http://aims.fao.org/aos/agrovoc/c_33261, http://aims.fao.org/aos/agrovoc/c_6377, http://aims.fao.org/aos/agrovoc/c_1513, |
Online Access: | http://agritrop.cirad.fr/389622/ http://agritrop.cirad.fr/389622/1/389622.pdf |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|