Proteolytic activities in Phaseolus vulgaris cotyledons under copper stress

Abstract

The changes in the protease activities of bean cotyledons were investigated in response to copper stress. Assays using synthetic substrates and specific protease inhibitors followed by activity measurements and electrophoresis analysis allowed to study the classes of enzymes involved in the storage protein mobilization during the germination of bean (Phaseolus vulgaris L) seeds, and then identify which ones were affected in the presence of 200 μM CuCl 2 in the imbibition medium. Copper treatment affected embryo growth and total protease activity. The results of SDS-gelatin-PAGE show that Cu excess led to a decrease in protease activity of 45 to 66 kDa. Moreover, cysteine-, aspartic-and metallo-protease activities were markedly lowered under copper stress, while serine-protease one was enhanced as well as its activity dependent abundance in comparison with control. However, the relative distribution of major cysteine protease in H 2 O-germinated seeds was significantly diminished after Cu exposure. Thus, copper excess can disturb the nitrogen freeing from reserve tissues at enzymatic level; differential responses of protease classes are discussed, notably, cysteine protease in the way of storage protein mobilization and serine protease in protective mechanism one.

Alleviation of copper toxicity in germinating pea seeds by IAA, GA3, Ca and citric acid

“…Subsequent studies have already shown that Cu imposed a delay in the embryonic axes growth of legume seeds (Sfaxi-Bousbih et al 2010;Karmous et al 2012;Chaoui and El Ferjani 2013). In the present experiment, a similar decrease in root and shoot length and fresh biomass in pea seeds during germination was found.…”

Ameliorative role of β-estradiol against lead-induced oxidative stressand genotoxic damage in germinating wheat seedlings

“…On the other hand, there are many studies reporting inhibitory effects of metals on amylase activity. Karmous et al (2012) found that cadmium and copper inhibited amylase activity in cotyledons of germinating seeds of numerous legumes. The inhibiting effect of lead on amylase activity might have resulted from the directly toxic effect of lead ions by displacing Ca 2+ ions.…”

Inhibition Of Proteolysis In Plant And Animal Tissues

“…Семена фасоли (Phaseolus vulgaris L.) проращивали в присутствии H 2 O и семядоли собирали через 6 дней согласно Karmous et al (2012). Семядоли гомогенизировали в ступке при 4 °С в четыре раза превышающем свежую массу объеме воды.…”

“…Определение содержания ингибиторов протеаз (α1-антипротеазного ингибитора и α2-макроглобулина) проводили по методу, основанному на БАПНА-амидазной реакции при значениях рН инкубационных сред 3,0; 3,8; 6,1; 7,2; 8,0 и 9,0. Cодержание определяемых ингибиторов протеаз выражали в г/кг свежей ткани (Karmous et al, 2012;Чиркин и др., 2012). Представлял интерес поиск общих мишеней для действия антиметаболита этионина на протеолитические ферменты человека и модельного организма.…”