0000000000485140
AUTHOR
M. Aulikki Salmia
Localization and Activity of a Carboxypeptidase in Germinating Seeds of Scots Pine, Pinus sylvestris
Extracts prepared from the endosperm of germinating seeds of Scots pine, Pinus sylvestris L., hydrolysed two typical carboxypeptidase substrates, Z-Phe-Ala and Z-Phe-Phe, with pH optima at 4.2 and 5.0. The activities were completely destroyed by diisopropylfluorophosphate. Identical heat inactivation curves and elution patterns in gel chromatography on Sephadex G-200 suggest that the two activities are due to a single enzyme. In resting seeds very low carboxypeptidase activity was present in both the endosperm and the embryo. During germination on agar gel at 20°C in the dark the activities, expressed as enzyme units per seed, increased in the seedling and particularly in the endosperm up t…
Activities of Two Peptidases in Resting and Germinating Seeds of Scots Pine, Pinus sylvestris
Extracts prepared from resting seeds of Scots pine, Pinus sylvestris L., rapidly hydrolysed two peptides, Leu–Tyr and Ala–Gly, at pH 8.6 and 7.8, respectively. In gel chromatography on Sephadex G-100 the two activities were eluted as separate peaks, which indicates that they are due to two different peptidases. The seeds were allowed to germinate at 20°C, the activities of the two enzymes were assayed separately on extracts from the endosperm and seedling tissues at different stages of germination, and compared with corresponding changes in dry weight and total nitrogen. Both enzyme activities were relatively high in the endosperm of resting seeds, and they increased about 2- and 3-fold dur…
Characterization of the Proteinases Present in Germinating Seeds of Scots Pine, Pinus sylvestris
Methods were developed to determine proteinase activity in germinating seeds of Scots pine. The assays were based on the liberation of TCA-soluble peptides from haemoglobin at pH 3.7 and from casein at pH 5.4 and pH 7.0; the reaction products were determined by the Lowry method. — Endosperms separated from seeds at the time of rapid storage protein mobilization (seedling length between 20 and 50 mm) showed high proteinase activities in all three assays. Experiments with different inhibitors suggested that at least four enzymes were involved. One of the enzymes resembled mammalian and microbial pepsin-like acid proteinases: the pH optimum was 3.7 and the enzyme was inhibited by pepstatin.—Th…
Localization and Activity of Naphthylamidases in Germinating Seeds of Scots Pine, Pinus sylvestris
Extracts prepared from endosperms of germinating seeds of Scots pine, Pinus sylvestris L., rapidly hydrolysed the β-naphthylamides of L-phenylalanine and L-leucine optimally at pH 6.5 and that of L-arginine at pH 7.7. Disc electrophoresis followed by activity staining showed that the activities were due to two naphthylamidases (aminopeptidases) with different substrate specificities. Seeds were allowed to germinate at 20°C on agar gel in the dark and the activities on the three substrates were assayed from separated endosperms and seedlings at various stages of germination. The activities in the endosperm of resting seeds were relatively high and they remained unchanged throughout the perio…
Inhibitors of endogenous proteinases in the seeds of Scots pine, Pinus sylvestris
Extracts of resting pine seeds inhibited the proteinase activities present in extracts of endosperms of germinating seeds (hydrolysis of haemoglobin at pH 3.7 and hydrolysis of casein at pH 5.4 and 7.0). Heating the extracts of resting seeds at 60°C destroyed their own proteinase activity but their proteinase inhibitor activity decreased by only 25 to 30%. Some properties of the inhibitor(s) were studied using extracts treated at 60°C. The inhibitor activities were non-dialysable. the inhibition increased linearly with increasing inhibitor concentration up to 80% of total proteinase activity, and the maximal inhibition was 80% at pH 3.7. 90% at pH 5.4. and 97% at pH 7.0. The extracts of res…