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81.
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84.
The previously reported class of potent inorganic inhibitors of Na,K-ATPase, named MCS factors, was shown to inhibit not only
Na,K-ATPase but several P-type ATPases with high potency in the sub-micromolar range. These MCS factors were found to bind
to the intracellular side of the Na, K-ATPase. The inhibition is not competitive with ouabain binding, thus excluding its
role as cardiac-steroid-like inhibitor of the Na,K-ATPase. The mechanism of inhibition of Na,K-ATPase was investigated with
the fluorescent styryl dye RH421, a dye known to report changes of local electric fields in the membrane dielectric. MCS factors
interact with the Na,K-ATPase in the E1 conformation of the ion pump and induce a conformational rearrangement that causes a change of the equilibrium dissociation
constant for one of the first two intracellular cation binding sites. The MCS-inhibited state was found to have bound one
cation (H+, Na+ or K+) in one of the two unspecific binding sites, and at high Na+ concentrations another Na+ ion was bound to the highly Na+-selective ion-binding site. 相似文献
85.
Testing for endocytosis in plants 总被引:6,自引:0,他引:6
Summary. For many years endocytosis has been regarded with great scepsis by plant physiologists. Although now generally accepted, care
must still be taken with experiments designed to demonstrate endocytic uptake at the plasma membrane. We have taken a critical
look at the various agents which are in use as markers for plant endocytosis, pointing out pitfalls and precautions which
should be taken. We also take this opportunity to introduce the tyrphostins – tyrosine kinase inhibitors –, which also seem
to prevent endocytosis in plants.
Correspondence and reprints: Heidelberg Institute for Plant Sciences, University of Heidelberg, Im Neuenheimer Feld 230, 69120
Heidelberg, Federal Republic of Germany. 相似文献
86.
A stilbene dye (Direct Yellow 11) and a methine dye, Basazol 46L, recalcitrant to common chemical
bleaches, were treated with horseradish and soybean peroxidases. Both enzymes were effective at chromophore
removal. When compared to laccase in combination with a mediator (ABTS), soybean peroxidase was more
effective at oxidative dye removal, especially for the methine dye. 相似文献
87.
In this paper we report an innovative and unconventional method based on circular dichroism for the identification of peanut DNA in food, which can be detected after PCR amplification at the nanomolar level by using an achiral PNA probe complementary to a tract of the peanut Ara h 2 gene and an achiral 3,3'-diethylthiadicarbocyanine dye [DiSC(2)(5)]. Peanuts are one of the most common causes of severe allergic reactions to foods and are particularly dangerous when they are "hidden" (undeclared) in food. For better protection of consumers, detection methods are required to specifically detect the presence of hidden allergens in a wide variety of food items. Alternative to the detection of the proteins is the determination of species-specific DNA, which is more resistant to technological treatments. PNAs are very specific probes able to recognize DNA sequences with high affinity and evidence for the binding can be obtained by using the DiSC(2)(5) dye, which aggregates onto the PNA-DNA duplex giving rise to a characteristic visibile band at 540 nm. Because the PNA-DNA duplex is in a right-handed helical conformation, the aggregation of the dye to the duplex gives also rise to a strong CD signal in the 500-600 nm region with a strong exciton coupling due to the formation of multimeric species, since the handedness of the helix is transferred to the dye aggregate. The dye does not interact with the free single-stranded DNA and although aggregating on the achiral PNA, this interaction is obviously not detectable by circular dichroism. Thus, only the formation of the PNA-DNA duplex, which takes place only upon specific Watson-Crick hydrogen binding between the PNA and the DNA bases, is detected, ensuring a very high specificity and sensitivity. The method has been optimized in a model system by using a synthetic oligonucleotide complementary to the PNA probe, showing that the intensity of the signal is linearly related to the amount of the DNA. The optimized method has been applied to the identification and quantitation of DNA extracted and amplified by PCR from peanuts and from peanut-containing foods, allowing for a very sensitive detection at a very low level (few pmol). 相似文献
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89.
Massimo Micaroni Giuseppe Perinetti Daniele Di Giandomenico Alexander Spaar 《Experimental cell research》2010,316(13):2071-2086
The mechanisms of secretory transport through the Golgi apparatus remain an issue of debate. The precise functional importance of calcium ions (Ca2+) for intra-Golgi transport has also been poorly studied. Here, using different approaches to measure free Ca2+ concentrations in the cell cytosol ([Ca2+]cyt) and inside the lumen of the Golgi apparatus ([Ca2+]GA), we have revealed transient increases in [Ca2+]cyt during the late phase of intra-Golgi transport that are concomitant with a decline in the maximal [Ca2+]GA restoration ability. Thus, this redistribution of Ca2+ from the Golgi apparatus into the cytosol during the movement of cargo through the Golgi apparatus appears to have a role in intra-Golgi transport, and mainly in the late Ca2+-dependent phase of SNARE-regulated fusion of Golgi compartments. 相似文献
90.
Eichlerová I Homolka L Nerud F 《Journal of industrial microbiology & biotechnology》2006,33(9):759-766
The little studied white rot fungus Ischnoderma resinosum was tested for its ability to decolorize seven different synthetic dyes. The strain efficiently decolorized Orange G, Amaranth, Remazol Brilliant Blue R, Cu-phthalocyanin and Poly R-478 on agar plates and in liquid culture at a relatively high concentration of 2–4 and 0.5–1 g l−1, respectively. Malachite Green and Crystal Violet were decolorized to a lower extent up to the concentration of 0.1 g l−1. Decolorization capacity of I. resinosum was higher than that in Phanerochaete chrysosporium, Pleurotus ostreatus or Trametes versicolor. In contrast with these thoroughly examined fungi, I. resinosum was able to degrade a wide spectrum of chemically and structurally different synthetic dyes. I. resinosum also efficiently decolorized dye mixtures. In liquid culture, Orange G and Remazol Brilliant Blue R were decolorized most rapidly; the process was not affected by different nitrogen content in the media. Shaken cultivation strongly inhibited the decolorization of Orange G. 相似文献