Rapid purification of polyamine oxidase from oat primary leaves

In vitro and in vivo inhibition of plant polyamine oxidase activity by polyamine analogues

“…66 kDa both in the DEAE-Sephacel and the gel filtration fractions. These results are similar to those reported by Federico et al (1989) and Li (1993). This protocol for purification of oat PAO was initially applied to low amounts of protein (Table 1), and further adjusted for the higher protein amounts used in the experiments described in this work.…”

“…This procedure was demonstrated to reduce PAO contamination with cytoplasmic proteins and also to prevent the dilution of PAO activity (Li, 1993). The precipitation caused by the addition of cold Me 2 CO, ion exchange, and gel filtration chromatographic steps completed the purification protocol used, yielding a 22.7 fold purification and a specific activity of 2181 nkat/mg (Table 1).…”

“…Extraction of apoplastic protein was performed as described by Li (1993) with slight modifications. The abaxial epidermal sheet of oat leaves was peeled and leaves were washed in distilled H 2 O.…”

Polyamine oxidase activity contributes to sustain maize leaf elongation under saline stress

“…Extraction of apoplastic proteins was performed as described by Li (1993) with slight modifications (Maiale et al, 2008). Plant material (40 g) was cut in 2 mm pieces, washed in distilled water and vacuum-infiltrated with 100 ml 5 mM potassium phosphate pH 6.5 added with 200 mM NaCl.…”

“…Table 1. Effects of inhibitors on apoplastic ROS-producing enzymes PAO was extracted and purified from apoplastic fluids of leaf blade segments according to Li (1993) with some modifications. PAO activity was determined according to Cona et al (2006b).…”