ArticleProceedings of the National Academy of Sciences of the United States of America1993
Expression of the functional mature chloroplast triose phosphate translocator in yeast internal membranes and purification of the histidine-tagged protein by a single metal-affinity chromatography step.
Article in Proceedings of the National Academy of Sciences of the United States of America, 1993. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed.
- Photosynthetic Electron Flows and Networks of Metabolite Trafficking to Sustain Metabolism in Photosynthetic Systems.Plants (Basel, Switzerland) · 2024Review
- Chlamydomonas mutants lacking chloroplast TRIOSE PHOSPHATE TRANSPORTER3 are metabolically compromised and light sensitive.The Plant cell · 2023Article
- The import and export business in plastids: transport processes across the inner envelope membrane.Plant physiology · 2011Article
- Molecular identification and functional characterization of Arabidopsis thaliana mitochondrial and chloroplastic NAD+ carrier proteins.The Journal of biological chemistry · 2009Article
- The plastidic bile acid transporter 5 is required for the biosynthesis of methionine-derived glucosinolates in Arabidopsis thaliana.The Plant cell · 2009Article
- Arabidopsis SAMT1 defines a plastid transporter regulating plastid biogenesis and plant development.The Plant cell · 2006Article
- The Arabidopsis plastidic glucose 6-phosphate/phosphate translocator GPT1 is essential for pollen maturation and embryo sac development.The Plant cell · 2005Article
- MIF signal transduction initiated by binding to CD74.The Journal of experimental medicine · 2003Article
- The plastidic pentose phosphate translocator represents a link between the cytosolic and the plastidic pentose phosphate pathways in plants.Plant physiology · 2002Article
- The phosphoenolpyruvate/phosphate translocator is required for phenolic metabolism, palisade cell development, and plastid-dependent nuclear gene expression.The Plant cell · 1999Article
- Molecular characterization of a carbon transporter in plastids from heterotrophic tissues: the glucose 6-phosphate/phosphate antiporter.The Plant cell · 1998Article
- A new class of plastidic phosphate translocators: a putative link between primary and secondary metabolism by the phosphoenolpyruvate/phosphate antiporter.The Plant cell · 1997Article
- Crystallization and identification of an assembly defect of recombinant antenna complexes produced in transgenic tobacco plants.Proceedings of the National Academy of Sciences of the United States of America · 1996Article
- Purification of a rat neurotensin receptor expressed in Escherichia coli.The Biochemical journal · 1996Article
- Engineering the lac permease for purification and crystallization.Journal of bioenergetics and biomembranes · 1996Review
- Isolation, characterization and expression of cDNA clones encoding a mitochondrial malate translocator from Panicum miliaceum L.Plant molecular biology · 1996Article
- Article
- Purification of Rhizobium leguminosarum HypB, a nickel-binding protein required for hydrogenase synthesis.Journal of bacteriology · 1994Article
- The 24 kDa outer envelope membrane protein from spinach chloroplasts: molecular cloning, in vivo expression and import pathway of a protein with unusual properties.Plant molecular biology · 1994Article
- Expression of genes encoding the tobacco chloroplast phosphate translocator is not light-regulated and is repressed by sucrose.Molecular & general genetics : MGG · 1994Article
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The mature part of the chloroplast triose phosphate-phosphate translocator was cloned into the yeast expression vector pEVP11. This construct was used to transform cells from both Saccharomyces cerevisiae and the fission yeast Schizosaccharomyces pombe. The chloroplast translocator protein was functionally expressed in the transformed yeast cells and represented about 1-2% of the Sch. pombe cell membrane protein. It was localized to mitochondrial membranes and/or membranes of the rough endoplasmic reticulum. In order to purify the recombinant translocator protein, a sequence encoding a C-terminal tag of six histidine residues was introduced into the corresponding cDNA. The expressed histidine-tagged translocator protein was purified from the transformed yeast cells under nondenaturing conditions to apparent homogeneity by a single-step affinity chromatography using a Ni2+. nitrilotriacetic acid resin. Both the expressed triose phosphate translocator and the recombinant histidine-tagged protein possess substrate specificities identical to those of the authentic chloroplast protein, providing definitive evidence for its identity as the triose phosphate translocator and further disproving its assignment as the receptor for chloroplast protein import. The yeast expression system in combination with the Ni2+. nitrilotriacetic acid chromatography thus provides a valuable tool for the production of purified membrane proteins in a functional state.
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