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  • 1
    ISSN: 1432-2048
    Keywords: Amino acid transport ; Carrier (amino acids) ; Plasma membrane ; Ricinus (amino acid transport) ; Root (amino acid transport)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The mechanism and specificity of amino-acid transport at the plasma membrane of Ricinus communis L. roots was investigated using membrane vesicles isolated by phase partitioning. The transport of glutamine, isoleucine, glutamic acid and aspartic acid was driven by both a pH gradient and a membrane potential (internally alkaline and negative), created artificially across the plasma membrane. This is consistent with transport via a proton symport. In contrast, the transport of the basic amino acids, lysine and arginine, was driven by a negative internal membrane potential but not by a pH gradient, suggesting that these amino acids may be taken up via a voltage-driven uniport. The energized uptake of all of the amino acids tested showed a saturable phase, consistent with carrier-mediated transport. In addition, the membrane-potential-driven transport of all the amino acids was greater at pH 5.5 than at pH 7.5, which suggests that there could be a direct pH effect on the carrier. Several amino-acid carriers could be resolved, based on competition studies: a carrier with a high affinity for a range of neutral amino acids (apart from asparagine) but with a low affinity for basic and acidic amino acids; a carrier which has a high affinity for a range of neutral amino acids except isoleucine and valine, but with a low affinity for basic and acidic amino acids; and a carrier which has a higher affinity for basic and some neutral amino acids but has a lower affinity for acidic amino acids. The existence of a separate carrier for acidic amino acids is discussed.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2048
    Keywords: Acetate uptake ; Beta-Glutamine transport ; Plasma membrane ; Ricinus ; Sucrose transport ; Tetraphenylphosphonium uptake
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Plasma membrane vesicles, purified by aqueous two-phase partitioning, were used to investigate the presence of sugar and amino acid carriers in cotyledons and roots of Ricinus communis L. and in roots of red beet (Beta vulgaris L.). Artificial pH and electrical gradients were generated across the plasma membrane, and [14C]acetate and [14C]tetraphenylphosphonium were used to demonstrate the presence of an internal alkaline pH gradient and an internal negative membrane potential, respectively. In Ricinus cotyledons, uptake of sucrose was more strongly inhibited than that of glutamine by p-chloromercuribenzenesulphonic acid, phlorizin and phenylglyoxal. The sucrose transport system showed a high degree of substrate specificity with only the presence of maltose and phenyl-α-glucoside significantly affecting sucrose uptake; in contrast, the glutamine transport system was inhibited by a number of other amino acids. ΔpH+gDψ-driven glutamine uptake showed saturation kinetics with a K m of 0.35 mol · m−3. Sucrose and glutamine Δψ-driven uptake was pH dependent with an optimum in the acidic range (pH 6.25) and a decrease at higher pH values. Vesicles obtained from cotyledons and roots of Ricinus showed different transport properties. In the cotyledons, gDH+gDψ-driven transport for both sucrose and glutamine were observed at similar levels; however, in the root tissue, δpH-Δψ-driven glutamine transport was the dominant uptake process. Uptake rates for glucose and fructose were low in the cotyledons whereas, in the roots, glucose and sucrose transport were slightly higher than that of fructose. In vesicles from red beet tissue there was a different uptake profile, with evidence of proton-coupled cotransport systems for sucrose and glucose, but lower uptake of glutamine and fructose. The results are discussed in relation to the reported different pathways for loading and unloading of solutes in these tissues.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2048
    Keywords: Glutamine transport ; Plasma membrane ; Proton cotransport ; Ricinus (solute transport) ; Sucrose transport ; Valinomycin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Evidence is presented for the proton-coupled transport of sucrose and glutamine in purified plasma membrane vesicles isolated from cotyledons ofRicinus communis. Imposition of a pH gradient (internal alkaline) across the plasma membrane resulted in a rapid uptake of sucrose and glutamine which was inhibited in the presence of carbonyl cyanide-m-chlorophenyl hydrazone. Imposition of a pH gradient plus an internal negative membrane potential stimulated uptake further. Glucose and fructose uptakes were negligible under these conditions. Sucrose uptake into the vesicles demonstrated saturation kinetics with a Km of 0.87 mol·m-3, indicating carrier-mediated transport. In support of this, uptake was very sensitive to the protein-modifying reagentp-chloromercuribenzenesulphonic acid. N-Ethylmaleimide, another sulphydryl reagent, was only slightly inhibitory. However, both reagents strongly inhibited sucrose uptake into intact cotyledons; the possible reasons for the difference between the intact and isolated systems are assessed. The value of this system for the study of sucrose and amino acid carriers is discussed.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2048
    Keywords: ATPase ; Plasma membrane ; Pyrophosphatase ; Ricinus (solute transport) ; Sucrose transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A highly enriched plasma membrane fraction has been isolated from dark-grown cotyledons ofRicinus communis by phase partitioning. This is demonstrated by the properties of the associated ATPase: high vanadate sensitivity, azide and nitrate insensitivity, sharp pH optimum around 6.5, and high specificity for ATP as substrate. The upper plasma membrane fraction also contained a pyrophosphatase activity, normally considered to be located on the tonoplast or Golgi membranes, which showed a specific activity higher than that in the lower phase. Sucrose gradient centrifugation of both microsomal and upper phase fractions showed a comigration of some pyrophosphatase activity with the plasma membrane fraction. Sucrose uptake changes with development inRicinus cotyledons. The ATPase activity in the upper (plasma membrane) phase also varied in a similar way with development, whereas activity in the lower phase showed little change. Pyrophosphatase activity in the upper phase also increased with development but did not show a peak and fall as seen for sucrose uptake and ATPase. The possibility that changes in plasma membrane ATPase may contribute to changes in sucrose uptake capacity and the possible cellular origin and physiological significance of the pyrophosphatase activity are discussed.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 165 (1991), S. 27-36 
    ISSN: 1615-6102
    Keywords: ATPase ; Cerium ; Cytochemistry ; Fixation ; Zea mays roots ; Plasma membrane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The cytochemical localization of ATPase activity has been investigated in maize root cells using both lead and cerium-based capture methods. With both methods, staining at the plasma membrane was observed in all cells of the root, although the precipitate obtained with cerium was more uniform and granular than that with lead. Controls using no substrate or no magnesium, β-glycerophosphate to replace ATP, vanadate or boiled tissue generally showed little or no staining. However, biochemical studies on purified plasma membrane fractions showed that ATPase activity was markedly inhibited by fixation, particularly by glutaraldehyde, and also by lead and cerium ions. Non-enzymic hydrolysis of ATP by cerium was greater than that by lead. The value and limitations of these procedures for the localization of plasma membrane H+-ATPase activity are summarized in relation to previous criticisms of these methods.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1615-6102
    Keywords: Analytical electron microscopy ; ATPase cytochemistry ; Plasma membrane ; Ricinus communis ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The claim that osmium-containing deposits which lack lead are frequently and incorrectly interpreted as enzymatic reaction products in lead precipitation techniques for ATPase localization in plants is without foundation. Proper controls clearly demonstrate the enzymatic origin of membrane-located deposits and the presence of lead is confirmed by analytical electron microscopy.
    Type of Medium: Electronic Resource
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