Affinity purification of sucrose binding proteins from the plant plasma membrane
| Autor: | Ze-Sheng Li, Andrée Bourbouloux, Serge Delrot, Abdel Majid Noubhani |
|---|---|
| Rok vydání: | 1994 |
| Předmět: |
Sucrose
Chromatography Elution Chemistry Cell Membrane Biophysics Membrane Proteins Biological Transport Maltose Ligand (biochemistry) Sucrose transport Biochemistry Chromatography Affinity Molecular Weight Sepharose chemistry.chemical_compound Membrane Affinity chromatography Structural Biology Genetics Plant Proteins |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression. 1219:389-397 |
| ISSN: | 0167-4781 |
| DOI: | 10.1016/0167-4781(94)90063-9 |
| Popis: | Purified plasma membranes from sugar beet leaves were solubilized by 1% 3-((3-cholamidopropyl)dimethylammonio)-1-propanesulfonate and loaded on a sepharose 6 B column substituted with sucrose. Elution with sucrose at pH 5.2 yielded a peak that represented 0.2% of the loaded protein. This peak did not appear when the samples were pretreated with either 0.5 mM N-ethylmaleimide (NEM) or 0.5 mM para-chloromercuribenzenesulfonic acid. It was also absent when palatinose, a sucrose analogue not recognized by the sucrose transporter, was used as the affinity ligand. The peak specifically eluted by sucrose from the sucrose-Sepharose column exhibited sucrose transport activity after reconstitution into proteoliposomes. This peak was further fractionated by ion-exchange chromatography on a Mono-Q column, and the different fractions obtained were differentially labeled by [3H]NEM in the presence of sugars recognized (sucrose, maltose) or not recognized (palatinose) by the sucrose transporter. The data allowed to identify two fractions that were enriched with two polypeptides (56 and 41 kDa) differentially labeled by NEM in the presence of sucrose. |
| Databáze: | OpenAIRE |
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