Cytosolic V-1-ATPase complex in pea (Pistum sativum L.) root cells was characterized by immunoblotting, immuno-electron microscopy and measurement of ATP hydrolysis activity. Using antibodies against the subunits A an...Cytosolic V-1-ATPase complex in pea (Pistum sativum L.) root cells was characterized by immunoblotting, immuno-electron microscopy and measurement of ATP hydrolysis activity. Using antibodies against the subunits A and B of V-type H+-ATPase (V-ATPase) from mung bean as primary antibodies, both immuno-blotting and immuno-electron microscopy showed that subunit A and subunit B existed in the cytosol. Measurement of ATPase activity further showed that cytosolic proteins had NO3--sensitive ATP hydrolysis activity. All these suggested that V-1-ATPase complex occurred in the cytosol. This is the direct proof that V-1-ATPase occurs in the cytosol of the plant cells for the first time.展开更多
Starch degradation in cells is closely associated with cereal seed germination, photosynthesis in leaves, carbohydrate storage in tuber and tuberous roots, and fleshy fruit development. Based on previously reported ...Starch degradation in cells is closely associated with cereal seed germination, photosynthesis in leaves, carbohydrate storage in tuber and tuberous roots, and fleshy fruit development. Based on previously reported in vitro assays, β amylase is considered as one of the key enzymes catalyzing starch breakdown, but up to date its role in starch breakdown in living cells remains unclear because the enzyme was shown often extrachloroplastic in living cells. Recently we have shown for the first time that β_amylase is predominantly immuno_localized to plastids in living cells of developing apple fruit. But it remains to know whether this model of β_amylase compartmentation is more widespread in plant living cells. The present experiment, conducted in tuberous root of sweet potato ( Ipomea batatas Lam. cv. Xushu 18) and via immunogold electron_microscopy technique, showed that β amylase visualized by gold particles was predominantly localized in plastids especially at periphery of starch granules, but the gold particles were scarcely found in other subcellular compartments, indicating that the enzyme is subcellularly compartmented in the same zone as its starch substrates. The density of gold particles (β amylase) in plastids was increasing during growing season, but the predominantly plastid_distributed pattern of β amylase in cells was shown unchanged throughout the tuberous root development. These data prove that the enzyme is compartmented in its functional sites, and so provide evidence to support the possible widespread biological function of the enzyme in catalyzing starch breakdown in plant living cells or at least in living cells of plant storage organs.展开更多
文摘Cytosolic V-1-ATPase complex in pea (Pistum sativum L.) root cells was characterized by immunoblotting, immuno-electron microscopy and measurement of ATP hydrolysis activity. Using antibodies against the subunits A and B of V-type H+-ATPase (V-ATPase) from mung bean as primary antibodies, both immuno-blotting and immuno-electron microscopy showed that subunit A and subunit B existed in the cytosol. Measurement of ATPase activity further showed that cytosolic proteins had NO3--sensitive ATP hydrolysis activity. All these suggested that V-1-ATPase complex occurred in the cytosol. This is the direct proof that V-1-ATPase occurs in the cytosol of the plant cells for the first time.
文摘Starch degradation in cells is closely associated with cereal seed germination, photosynthesis in leaves, carbohydrate storage in tuber and tuberous roots, and fleshy fruit development. Based on previously reported in vitro assays, β amylase is considered as one of the key enzymes catalyzing starch breakdown, but up to date its role in starch breakdown in living cells remains unclear because the enzyme was shown often extrachloroplastic in living cells. Recently we have shown for the first time that β_amylase is predominantly immuno_localized to plastids in living cells of developing apple fruit. But it remains to know whether this model of β_amylase compartmentation is more widespread in plant living cells. The present experiment, conducted in tuberous root of sweet potato ( Ipomea batatas Lam. cv. Xushu 18) and via immunogold electron_microscopy technique, showed that β amylase visualized by gold particles was predominantly localized in plastids especially at periphery of starch granules, but the gold particles were scarcely found in other subcellular compartments, indicating that the enzyme is subcellularly compartmented in the same zone as its starch substrates. The density of gold particles (β amylase) in plastids was increasing during growing season, but the predominantly plastid_distributed pattern of β amylase in cells was shown unchanged throughout the tuberous root development. These data prove that the enzyme is compartmented in its functional sites, and so provide evidence to support the possible widespread biological function of the enzyme in catalyzing starch breakdown in plant living cells or at least in living cells of plant storage organs.
