Journal:Acta Cryst D:S2059798325007089
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<b>Molecular Tour</b><br> | <b>Molecular Tour</b><br> | ||
| - | When cells build and maintain their membranes, they need a balance of protein insertion, folding, and degradation. In Escherichia coli, this process is hypothetic to involve the AAA+ protease FtsH, the insertase YidC, and the regulatory HflKC complex. Their interaction had not been shown at structural level. To test this idea, we used single-particle cryo-electron microscopy on detergent-solubilized samples enriched for these proteins. | + | When cells build and maintain their membranes, they need a balance of protein insertion, folding, and degradation. In ''Escherichia coli'', this process is hypothetic to involve the AAA+ protease FtsH, the insertase YidC, and the regulatory HflKC complex. Their interaction had not been shown at structural level. To test this idea, we used single-particle cryo-electron microscopy on detergent-solubilized samples enriched for these proteins. |
The results were unexpected. Instead of clear views of an FtsH–HflKC–YidC assembly, the datasets revealed high-resolution structures of ArnA, an enzyme linked to polymyxin resistance, and AcrB, the multidrug efflux transporter of the AcrAB–TolC system. Both proteins are known to appear during affinity purification, but their repeated presence across different methods and even in membrane fractions suggests that their recovery was not purely accidental. ArnA, usually described as cytoplasmic, was consistently found in membrane-enriched samples, while AcrB is a well-established membrane protein. We also observed class averages resembling GroEL and cytochrome bo3 oxidase. | The results were unexpected. Instead of clear views of an FtsH–HflKC–YidC assembly, the datasets revealed high-resolution structures of ArnA, an enzyme linked to polymyxin resistance, and AcrB, the multidrug efflux transporter of the AcrAB–TolC system. Both proteins are known to appear during affinity purification, but their repeated presence across different methods and even in membrane fractions suggests that their recovery was not purely accidental. ArnA, usually described as cytoplasmic, was consistently found in membrane-enriched samples, while AcrB is a well-established membrane protein. We also observed class averages resembling GroEL and cytochrome bo3 oxidase. | ||
Revision as of 15:09, 26 August 2025
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