scholarly journals Effect of ABA and GA3 on protein mobilization in embryos and cotyledons of angico [Anadenanthera peregrina (L.) speg] seeds during germination

Author(s):  
Douglas Barduche ◽  
Renato Paiva ◽  
Mauricio A. Lopes ◽  
Edilson Paiva

In this work, a woody species [A. peregrina (L.) Speg.] was studied in order to observe the effect of ABA and GA3 at the biochemical level during the process of seed germination. Embryos incubated in sucrose solution containing ABA and/or GA3 were analyzed through SDS-PAGE to observe the mobilization pattern of storage proteins during the beginning of germination. Cotyledons isolated from seeds incubated in aqueous solutions containing ABA and/or GA3, were also analyzed through SDS-PAGE and by PAGE/Activity Gels (polyacrylamide gels copolymerized with substrate for enzymes) to observe the mobilization pattern of storage proteins and protease activity after the beginning of the germination. Results of these experiments show that ABA blocks protein mobilization by inhibiting protease activity in cotyledons. This inhibition is not sufficient to prevent germination showing that the effect of ABA on germination is not dependent on protease activity. The blockage of storage protein mobilization was also observed in embryos, but no protease activity inhibition was clearly detected. ABA was able to induce the synthesis of proteins in cotyledons but not in embryos. A polypeptide with an approximate molecular weight of 17 kD, was degraded within 6 hours in control embryos, but this degradation was blocked by ABA and GA3. Using the same concentrations of ABA and GA3 on embryos and cotyledons, the effect of ABA was counteracted by GA3 in embryos, but not in cotyledons. Although the effects of ABA and GA3 were not so different from those shown in the literature, the behavior of 17 kD-polypeptide contradicts these reports suggesting that specific studies should be performed.

2004 ◽  
Vol 51 (3) ◽  
pp. 609-624 ◽  
Author(s):  
Małgorzata Grudkowska ◽  
Barbara Zagdańska

Cysteine proteinases also referred to as thiol proteases play an essential role in plant growth and development but also in senescence and programmed cell death, in accumulation of storage proteins such as in seeds, but also in storage protein mobilization. Thus, they participate in both anabolic and catabolic processes. In addition, they are involved in signalling pathways and in the response to biotic and abiotic stresses. In this review an attempt was undertaken to illustrate these multiple roles of cysteine proteinases and the mechanisms underlying their action.


Parasitology ◽  
2007 ◽  
Vol 135 (1) ◽  
pp. 125-130 ◽  
Author(s):  
V. M. BOWLES ◽  
A. R. YOUNG ◽  
S. C. BARKER

SUMMARYTo investigate the biochemical components of egg-hatch in the body louse, Pediculus humanus, egg-shell-washings (ESW) were collected during the first 2 h post-hatching and analysed by gelatin SDS-PAGE. These ESW contained proteases with molecular mass in the range of 25–100 kDa; the most abundant proteases were ~25 kDa. The 3 main regions of protease activity in the one-dimensional gelatin SDS-PAGE gels resolved to at least 23 distinct regions of protease activity when analysed by two-dimensional gelatin SDS-PAGE, with iso-electric points spread over the entire 3 to 10 pH range. Mechanistic characterization indicated that the ESW contained proteases of the metallo-class, inhibited by both 1,10-phenanthroline and EDTA. Several protease inhibitors were tested for their ability to inhibit louse egg-hatch in vitro. The metalloprotease inhibitor 1,10-phenanthroline and the aminopeptidase inhibitor bestatin significantly inhibited (P<0·05) louse egg-hatch (100% and 58%, respectively). The presence of metalloproteases at the time of egg-hatch and the inhibition of egg-hatch in P. humanus by metalloprotease inhibitors suggests a crucial role for these proteases in the hatching of this medically important parasite.


2018 ◽  
Vol 14 (5-6) ◽  
Author(s):  
Abeer Alhendi ◽  
Wade Yang ◽  
Paul J. Sarnoski

AbstractInactivation of pure soybean lipoxygenase (LOX) by pulsed light (PL) technique was found to occur due to a photochemical effect, while inactivation of soybean LOX in a real food (soymilk) was due to the photothermal effect of PL. The effect of solution properties on the photochemical ability of PL to inactivate and degrade LOX was investigated. LOX was placed in different conditions and treated with PL at a 7 cm distance with different times. The result showed that LOX was less stable during PL operation at pH 9 compared with pH 6.8. Increasing LOX concentration, adding starch, and making a colored solution did reduce the photochemical ability of PL to inactivate LOX. PL and thermal treatment of partially purified LOX degraded the LOX band (measured by using SDS-PAGE) when no protease inhibitors were added. Controlling protease activity led to degradation of LOX by PL but not by thermal treatment.


1990 ◽  
Vol 272 (3) ◽  
pp. 677-682 ◽  
Author(s):  
M A Belozersky ◽  
Y E Dunaevsky ◽  
N E Voskoboynikova

A homogeneous preparation of metalloproteinase, purified 1000-fold, was obtained from buckwheat (Fagopyrum esculentum) seeds. The Mr of the enzyme, determined by SDS/PAGE, was 34,000 (it was 39,000 by gel chromatography). Its pH optimum was 8.0-8.2 with 13 S globulin, from buckwheat seeds, as substrate. Atomic-absorption spectroscopy revealed the presence of one Zn2+ ion per enzyme molecule. The enzyme was completely inhibited by EDTA (1 mM), zincone (1 mM) and 1, 10-phenanthroline (1 mM). The metalloproteinase performed limited proteolysis of the following seed storage proteins: 13 S globulin from buckwheat seeds and 11 S globulin from soybean (Glycine max) seeds. It hydrolysed three peptide bonds formed by the amino groups of Leu15, Tyr16 and Phe25 in the oxidized B-chain of insulin. In its main properties the enzyme is similar to metalloproteinases of animal and bacterial origin.


2004 ◽  
Vol 31 (1) ◽  
pp. 36-41 ◽  
Author(s):  
A. V. Agafonov ◽  
N. B. Sukhareva ◽  
S. O. Baturin ◽  
O. A. Struzhkina

Author(s):  
Ananya Panda ◽  
Swapan K. Tripathy

Total seed storage protein profiles of 74 mungbean land races, three wild accessions and a popular variety ‘Jyoti’ of Odisha were analysed by Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). 32 genotypes could be clearly identified based on genotype-specific seed protein fingerprints while rest of the test genotypes were categorized into eight protein types. Genotypes included in each protein type had 100% homology and some of these could be duplicates. In this pursuit, a few specific polypeptide markers have been detected for identification of the land races/ genotypes. Dendrogram based on electrophoretic data clustered the genotypes into seven groups at 70% phenon level. Paralakhemundi local, Samarjhola local and Phulbani local-D; and three wild accessions (TCR 20, TCR 213 and TCR 243) were comparatively divergent from other genotypes. Besides, Jyoti, Kalahandi local 2A, Sikri local, kodala local A and TCR 20 were identified to be protein rich with high seed yield. TCR 20 being morphologically similar to mungbean, moderately high protein content and high yielding as well as resistant to drought and bruchids; it may serve as a valuable source genotype in recombination breeding


2006 ◽  
Vol 52 (1) ◽  
pp. 16-23 ◽  
Author(s):  
José de Jesús Serrano-Luna ◽  
Isaac Cervantes-Sandoval ◽  
Jesús Calderón ◽  
Fernando Navarro-García ◽  
Victor Tsutsumi ◽  
...  

Acanthamoeba spp. are free-living amoebae that cause amoebic granulomatous encephalitis, skin lesions, and ocular amoebic keratitis in humans. Several authors have suggested that proteases could play a role in the pathogenesis of these diseases. In the present work, we performed a partial biochemical characterization of proteases in crude extracts of Acanthamoeba spp. and in conditioned medium using 7.5% SDS–PAGE copolymerized with 0.1% m/v gelatin as substrate. We distinguished a total of 17 bands with proteolytic activity distributed in two species of Acanthamoeba. The bands ranged from 30 to 188 kDa in A. castellanii and from 34 to 144 kDa in A. polyphaga. Additionally, we showed that the pattern of protease activity differed in the two species of Acanthamoeba when pH was altered. By using protease inhibitors, we found that the proteolytic activities belonged mostly to the serine protease family and secondly to cysteine proteases and that the proteolytic activities from A. castellanii were higher than those in A. polyphaga. Furthermore, aprotinin was found to in hibit crude extract protease activity on Madin–Darby canine kidney (MDCK) monolayers. These data suggest that protease patterns could be more complex than previously reported.Key words: Acanthamoeba spp., amoebic keratitis, serine proteases, cysteine proteases, cytopathic effect.


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