Isolation and characterization of a sucrose carrier cDNA from spinach by functional expression in yeast.

Riesmeier, Jörg W.; Willmitzer, Lothar; Frommer, Wolf B

doi:10.1002/j.1460-2075.1992.tb05575.x

Cited by 438 publications

(401 citation statements)

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“…The resulting plasmid pET1 (AATP1 cDNA under control of the T7 promoter) was subsequently linearized with Notl and again partially digested with EcoRI. For overexpression of AATP1 in yeast under the control of the ADH promoter, the 2.2 kb Notl/EcoRI fragment of pET1 was inserted into p112A1NE (Riesmeier et al, 1992) cut with Nod and EcoRl. The resulting plasmid was named pET12.…”

Section: Dna Constructions For Heterologous Expression Of Aatp1 Protementioning

confidence: 99%

Characterization of a novel eukaryotic ATP/ADP translocator located in the plastid envelope of Arabidopsis thaliana L.

et al. 1997

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SummaryRecently, we have sequenced a cDNA clone from Arabidopsis thaliana L. encoding a novel putative ATP/ADP translocator (AATP1). Here, we demonstrate that the radioactively labeled AATP1 precursor protein, synthesized in vitro, is targeted to envelope membranes of isolated spinach chloroplasts. Antibodies raised against a synthetic peptide of AATP1 recognized a single polypeptide of about 62 kDa in chloroplast inner envelope preparations. The cDNA coding for the AATP1 protein was functionally expressed in Saccharomyces cerevisiae and Escherichia coil In both expression systems, increased rates of ATP transport were observed after reconstitution of the extracted protein into proteoliposomes. To our knowledge, this is the first report on the functional expression of an intrinsic plant membrane protein in E. coil To yield high rates of ATP transport, proteoliposomes had to be preloaded with ADP, indicating a counter-exchange mode of transport. Carboxyatractyloside did not substantially interfere with ATP transport into proteoliposomes containing the plastidic ATP/ADP translocator. An apparent KM for ATP of 28 llM was determined which is similar to values reported for isolated plastidso The data presented here strongly support the conclusion that AATP1 represents a novel eukaryotic adenylate carrier and that it is identical with the so far unknown plastidic ATP/ADP translocator.

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Section: Dna Constructions For Heterologous Expression Of Aatp1 Protementioning

confidence: 99%

Characterization of a novel eukaryotic ATP/ADP translocator located in the plastid envelope of Arabidopsis thaliana L.

et al. 1997

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“…Sucrose molecules cross the plasma membrane of phloem cells through specific carriers. Cloning of the SUT1 gene encoding sucrose carrier by yeast complementation [1,2] was the basis for identifying sucrose carriers in other plant species [3 5]. Those carriers work as proton/sucrose cotransporters and have been localized to the phloem, either in companion cells or in sieve elements [6,7] However, the exit of sucrose from phloem cells and subsequent import into sink organs is less clearly understood.…”

Section: Introductionmentioning

confidence: 99%

Identification of a pollen‐specific sucrose transporter‐like proteinNtSUT3 from tobacco

Lemoine

Bürkle²,

Barker³

et al. 1999

FEBS Letters

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Pollen cells are symplasmieally isolated during maturation and germination. Pollen therefore needs to take up nutrients via membrane carriers. Physiological measurements on pollen indicate sucrose transport in the pollen tube. A cDNA encoding a pollen-specific sucrose transporter-like protein NtSUT3 was isolated from a tobacco pollen cDNA library. NtSUT3 expression is detected only in pollen and is restricted to late pollen development, pollen germination and pollen tube growth. Altogether these data indicate that pollen is supplied not only with glucose, but also with sucrose through a specific sucrose transporter. The respective contribution of each transport pathway may change during pollen tube growth.

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“…The movement of H+ and Suc is tightly coupled with a stoichiometry of 1:l (Slone and Buckhout, 1991), and the carrier is specific for the Suc molecule, although phenylglucosides are also recognized by the carrier (Hecht et al, 1992). Finally, a gene encoding the H+-Suc symport protein was recently identified in spinach leaf cells (Riesmeier et al, 1992). In the plant, this transporter is responsible for phloem loading and, thus, photoassimilate export from leaves in many plant species (Giaquinta, 1983; Bush, 1993; Riesmeier et al, 1993).…”

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Kinetics Analysis of the Plasma Membrane Sucrose-H+ Symporter from Sugar Beet (Beta vulgaris L.) Leaves

Buckhout¹

1994

Plant Physiol.

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The kinetics behavior of the H+-sucrose (Suc) symporter was investigated in plasma membrane vesicles from sugar beet (Beta vulgaris 1.) leaves by analyzing the effect of externa1 and internal pH (pH. and pHi, respectively) on Suc uptake. l h e apparent K,,, for Suc uptake increased 18-fold as the pH. increased from 5.5 to 7.5. Over this same pH, range, the apparent VmaX for Suc uptake remained constant. l h e effects of pHi in the presence or absence of internal Suc were exclusively restrided to changes in Vmx. Thus, proton concentration on the inside of the membrane vesicles ([H+]i) behaved as a noncompetitive inhibitor of Suc uptake. l h e K,,, for the proton concentration on the outside of the membrane vesicles was estimated to be pH 6.3, which would indicate that at physiological apoplastic pH Suc transport might be sensitive to changes in pH,. On the other hand, the Suc transport across the PM is catalyzed by an H+-Suc symporter (Buckhout, 1989;Bush, 1989; Lemoine and Delrot, 1989; Williams et al., 1990). Transport is driven by the proton motive force established across the PM by the H+-ATPase in vivo and is electrogenic (Bush, 1990; Slone and Buckhout, 1991). The movement of H+ and Suc is tightly coupled with a stoichiometry of 1:l (Slone and Buckhout, 1991), and the carrier is specific for the Suc molecule, although phenylglucosides are also recognized by the carrier (Hecht et al., 1992). Finally, a gene encoding the H+-Suc symport protein was recently identified in spinach leaf cells (Riesmeier et al., 1992). In the plant, this transporter is responsible for phloem loading and, thus, photoassimilate export from leaves in many plant species (Giaquinta, 1983;Bush, 1993;Riesmeier et al., 1993).The membrane vesicle system offers the advantage for the kinetics analysis of transport processes that the driver ion and solute concentration on both sides of the vesicle mem- brane can be varied, and it has the further advantage that effects of tissue thickness and substrate access are eliminated (Ehwald et al., 1979). Determination of the kinetics parameters of a transport process can provide predictive insight into the behavior of the transporter in vivo. For example, if the apparent K, for [H' ], of the H+-Suc symporter is approximately equal to pH,, then changes in pH, could significantly affect the rate of Suc uptake. The same would hold true for for inhibition of uptake by [H+]i. Thus, a decrease in pH, might significantly inhibit Suc uptake, as is the case for H+-dependent C1-uptake in Chara (Sanders and Hansen, 1981).On the other hand, a single carrier might be involved in influx and efflux depending on the local concentrations of the ligands and the relative sensitivity of the carrier to changes in ligand concentration. Such macroscopic reversibility is found, for example, in the Na+-HC03-symporter in glial cells. This symporter catalyzes the movement of Na+ and HC03-with inward transport being stimulated by increasing concentration of HC03-outside of the membrane vesicles or by membrane depolarization, ...

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Isolation and characterization of a sucrose carrier cDNA from spinach by functional expression in yeast.

Cited by 438 publications

References 36 publications

Characterization of a novel eukaryotic ATP/ADP translocator located in the plastid envelope of Arabidopsis thaliana L.

Characterization of a novel eukaryotic ATP/ADP translocator located in the plastid envelope of Arabidopsis thaliana L.

Identification of a pollen‐specific sucrose transporter‐like proteinNtSUT3 from tobacco

Kinetics Analysis of the Plasma Membrane Sucrose-H+ Symporter from Sugar Beet (Beta vulgaris L.) Leaves

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