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| | <StructureSection load='4jfb' size='340' side='right'caption='[[4jfb]], [[Resolution|resolution]] 3.80Å' scene=''> | | <StructureSection load='4jfb' size='340' side='right'caption='[[4jfb]], [[Resolution|resolution]] 3.80Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[4jfb]] is a 6 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4JFB OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4JFB FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[4jfb]] is a 6 chain structure with sequence from [https://en.wikipedia.org/wiki/Escherichia_coli_K-12 Escherichia coli K-12]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4JFB OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4JFB FirstGlance]. <br> |
| - | </td></tr><tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[2zfg|2zfg]], [[3k1g|3k1g]], [[3k19|3k19]]</td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 3.801Å</td></tr> |
| - | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4jfb FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4jfb OCA], [http://pdbe.org/4jfb PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4jfb RCSB], [http://www.ebi.ac.uk/pdbsum/4jfb PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4jfb ProSAT]</span></td></tr> | + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=4jfb FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4jfb OCA], [https://pdbe.org/4jfb PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4jfb RCSB], [https://www.ebi.ac.uk/pdbsum/4jfb PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4jfb ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/OMPF_ECOLI OMPF_ECOLI]] Forms pores that allow passive diffusion of small molecules across the outer membrane. It is also a receptor for the bacteriophage T2.<ref>PMID:19721064</ref> | + | [https://www.uniprot.org/uniprot/OMPF_ECOLI OMPF_ECOLI] Forms pores that allow passive diffusion of small molecules across the outer membrane. It is also a receptor for the bacteriophage T2.<ref>PMID:19721064</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| | + | [[Category: Escherichia coli K-12]] |
| | [[Category: Large Structures]] | | [[Category: Large Structures]] |
| - | [[Category: Chaptal, V]] | + | [[Category: Chaptal V]] |
| - | [[Category: Falson, P]] | + | [[Category: Falson P]] |
| - | [[Category: Jault, J M]] | + | [[Category: Jault JM]] |
| - | [[Category: Kilburg, A]] | + | [[Category: Kilburg A]] |
| - | [[Category: Reyes-Meija, G C]] | + | [[Category: Reyes-Meija GC]] |
| - | [[Category: Sarwan, J]] | + | [[Category: Sarwan J]] |
| - | [[Category: Wiseman, B]] | + | [[Category: Wiseman B]] |
| - | [[Category: E. coli outer membrane]]
| + | |
| - | [[Category: Membrane protein]]
| + | |
| - | [[Category: Ompf]]
| + | |
| - | [[Category: Porin]]
| + | |
| Structural highlights
Function
OMPF_ECOLI Forms pores that allow passive diffusion of small molecules across the outer membrane. It is also a receptor for the bacteriophage T2.[1]
Publication Abstract from PubMed
Despite the growing interest in membrane proteins, their crystallization remains a major challenge. In the course of a crystallographic study on the multidrug ATP-binding cassette transporter BmrA, mass spectral analyses on samples purified with six selected detergents revealed unexpected protein contamination visible for the most part on overloaded SDS-PAGE. A major contamination from the outer membrane protein OmpF was detected in purifications with Foscholine 12 (FC12) but not with Lauryldimethylamine-N-oxide (LDAO) or any of the maltose-based detergents. Consequently, in the FC12 purified BmrA, OmpF easily crystallized over BmrA in a new space group, and whose structure is reported here. We therefore devised an optimized protocol to eliminate OmpF during the FC12 purification of BmrA. On the other hand, an additional band visible at approximately 110 kDa was detected in all samples purified with the maltose-based detergents. It contained AcrB that crystallized over BmrA despite its trace amounts. Highly pure BmrA preparations could be obtained using either a DeltaacrAB E. coli strain and n-dodecyl-beta-D-maltopyranoside, or a classical E. coli strain and lauryl maltose neopentyl glycol for the overexpression and purification, respectively. Overall our results urge to incorporate a proteomics-based purity analysis into quality control checks prior to commencing crystallization assays of membrane proteins that are notoriously arduous to crystallize. Moreover, the strategies developed here to selectively eliminate obstinate contaminants should be applicable to the purification of other membrane proteins overexpressed in E. coli.
Stubborn contaminants: influence of detergents on the purity of the multidrug ABC transporter BmrA.,Wiseman B, Kilburg A, Chaptal V, Reyes-Mejia GC, Sarwan J, Falson P, Jault JM PLoS One. 2014 Dec 17;9(12):e114864. doi: 10.1371/journal.pone.0114864., eCollection 2014. PMID:25517996[2]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Duval V, Nicoloff H, Levy SB. Combined inactivation of lon and ycgE decreases multidrug susceptibility by reducing the amount of OmpF porin in Escherichia coli. Antimicrob Agents Chemother. 2009 Nov;53(11):4944-8. doi: 10.1128/AAC.00787-09., Epub 2009 Aug 31. PMID:19721064 doi:10.1128/AAC.00787-09
- ↑ Wiseman B, Kilburg A, Chaptal V, Reyes-Mejia GC, Sarwan J, Falson P, Jault JM. Stubborn contaminants: influence of detergents on the purity of the multidrug ABC transporter BmrA. PLoS One. 2014 Dec 17;9(12):e114864. doi: 10.1371/journal.pone.0114864., eCollection 2014. PMID:25517996 doi:http://dx.doi.org/10.1371/journal.pone.0114864
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