4DNE
Crystal structure of a triple-mutant of streptavidin in complex with desthiobiotin
Summary for 4DNE
| Entry DOI | 10.2210/pdb4dne/pdb |
| Descriptor | Streptavidin, 6-(5-METHYL-2-OXO-IMIDAZOLIDIN-4-YL)-HEXANOIC ACID, SULFATE ION, ... (4 entities in total) |
| Functional Keywords | biotin, biotin-binding protein |
| Biological source | Streptomyces avidinii |
| Cellular location | Secreted: P22629 |
| Total number of polymer chains | 2 |
| Total formula weight | 38788.46 |
| Authors | Panwar, P.,Deniaud, A.,Pebay-Peyroula, E. (deposition date: 2012-02-08, release date: 2012-09-26, Last modification date: 2023-09-13) |
| Primary citation | Panwar, P.,Deniaud, A.,Pebay-Peyroula, E. Contamination from an affinity column: an encounter with a new villain in the world of membrane-protein crystallization. Acta Crystallogr.,Sect.D, 68:1272-1277, 2012 Cited by PubMed Abstract: Attempts to crystallize AtNTT1, a chloroplast ATP/ADP transporter from Arabidopsis thaliana, revealed an unexpected contaminant, Strep-Tactin, a variant of streptavidin that was used during purification of the protein. Although it was present in very small amounts, crystals of Strep-Tactin were reproducibly grown from the AtNTT1 solution. AtNTT1 was overexpressed in Escherichia coli and purified from detergent-solubilized membrane fractions using Strep-Tactin affinity chromatography based on an engineered streptavidin. The contamination of protein solutions purified on Strep-Tactin columns has never been described previously and seems to be specific to membrane proteins solubilized in detergents. Trace amounts of Strep-Tactin were observed to be eluted from a Strep-Tactin column using several routinely used detergents, illustrating their possible role in the contamination. This finding raises an alarm and suggests caution in membrane-protein purification using Strep-Tactin affinity columns, where detergents are essential components. The small crystals of contaminant protein led to the structure at 1.9 Å resolution of Strep-Tactin in complex with desthiobiotin. PubMed: 22993081DOI: 10.1107/S090744491202639X PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.88 Å) |
Structure validation
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