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| - | [[Image:1g8e.gif|left|200px]]<br /><applet load="1g8e" size="450" color="white" frame="true" align="right" spinBox="true" | |
| - | caption="1g8e, resolution 1.8Å" /> | |
| - | '''CRYSTAL STRUCTURE OF FLHD FROM ESCHERICHIA COLI'''<br /> | |
| | | | |
| - | ==Overview== | + | ==CRYSTAL STRUCTURE OF FLHD FROM ESCHERICHIA COLI== |
| - | FlhD is a 13.3 kDa transcriptional activator protein of flagellar genes, and a global regulator. FlhD activates the transcription of class II, operons in the flagellar regulon when complexed with a second protein FlhC, (21.5 kDa). FlhD also regulates other expression systems in Escherichia, coli. We are seeking to understand this plasticity of FlhD's DNA-binding, specificity and, to this end, we have determined the crystal structure of, the isolated FlhD protein. The structure was solved by substituting, seleno-methionine for natural sulphur-methionine in FlhD, crystallizing, the protein and determining the structure factor phases by the method of, multiple-energy anomalous dispersion (MAD). The FlhD protein is dimeric., The dimer is tightly coupled, with an intimate contact surface, implying, that the dimer does not easily dissociate. The FlhD monomer is, predominantly alpha-helical. The C-termini of both FlhD monomers (residues, 83-116) are completely disrupted by crystal packing, implying that this, region of FlhD is highly flexible. However, part of the C-terminus, structure in chain A (residues 83-98) was modelled using a native FlhD, crystal. What is seen in chain A suggests a classic DNA-binding, helix-turn-helix (HTH) motif. FlhD does not bind DNA by itself, so it may, be that the DNA-binding HTH motif becomes rigidly defined only when FlhD, forms a complex with some other protein, such as FlhC. If this were true, it might explain how FlhD exhibits plasticity in its DNA-binding, specificity, as each partner protein with which it forms a complex could, allosterically affect the binding specificity of its HTH motif. A, disulphide bridge is seen between the unique cysteine residues (Cys-65) of, FlhD native homodimers. Alanine substitution at Cys-65 does not affect, FlhD transcription activator activity, suggesting that the disulphide bond, is not necessary for either dimer stability or this function of FlhD., Electrostatic potential analysis indicates that dimeric FlhD has a, negatively charged surface. | + | <StructureSection load='1g8e' size='340' side='right'caption='[[1g8e]], [[Resolution|resolution]] 1.80Å' scene=''> |
| | + | == Structural highlights == |
| | + | <table><tr><td colspan='2'>[[1g8e]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1G8E OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1G8E FirstGlance]. <br> |
| | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.8Å</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=1g8e FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1g8e OCA], [https://pdbe.org/1g8e PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1g8e RCSB], [https://www.ebi.ac.uk/pdbsum/1g8e PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1g8e ProSAT]</span></td></tr> |
| | + | </table> |
| | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/FLHD_ECOLI FLHD_ECOLI] Functions in complex with FlhC as a master transcriptional regulator that regulates transcription of several flagellar and non-flagellar operons by binding to their promoter region. Activates expression of class 2 flagellar genes, including fliA, which is a flagellum-specific sigma factor that turns on the class 3 genes. Also regulates genes whose products function in a variety of physiological pathways.<ref>PMID:7961507</ref> <ref>PMID:11169100</ref> <ref>PMID:15941987</ref> <ref>PMID:18765794</ref> |
| | + | == Evolutionary Conservation == |
| | + | [[Image:Consurf_key_small.gif|200px|right]] |
| | + | Check<jmol> |
| | + | <jmolCheckbox> |
| | + | <scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/g8/1g8e_consurf.spt"</scriptWhenChecked> |
| | + | <scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview03.spt</scriptWhenUnchecked> |
| | + | <text>to colour the structure by Evolutionary Conservation</text> |
| | + | </jmolCheckbox> |
| | + | </jmol>, as determined by [http://consurfdb.tau.ac.il/ ConSurfDB]. You may read the [[Conservation%2C_Evolutionary|explanation]] of the method and the full data available from [http://bental.tau.ac.il/new_ConSurfDB/main_output.php?pdb_ID=1g8e ConSurf]. |
| | + | <div style="clear:both"></div> |
| | + | <div style="background-color:#fffaf0;"> |
| | + | == Publication Abstract from PubMed == |
| | + | FlhD is a 13.3 kDa transcriptional activator protein of flagellar genes and a global regulator. FlhD activates the transcription of class II operons in the flagellar regulon when complexed with a second protein FlhC (21.5 kDa). FlhD also regulates other expression systems in Escherichia coli. We are seeking to understand this plasticity of FlhD's DNA-binding specificity and, to this end, we have determined the crystal structure of the isolated FlhD protein. The structure was solved by substituting seleno-methionine for natural sulphur-methionine in FlhD, crystallizing the protein and determining the structure factor phases by the method of multiple-energy anomalous dispersion (MAD). The FlhD protein is dimeric. The dimer is tightly coupled, with an intimate contact surface, implying that the dimer does not easily dissociate. The FlhD monomer is predominantly alpha-helical. The C-termini of both FlhD monomers (residues 83-116) are completely disrupted by crystal packing, implying that this region of FlhD is highly flexible. However, part of the C-terminus structure in chain A (residues 83-98) was modelled using a native FlhD crystal. What is seen in chain A suggests a classic DNA-binding, helix-turn-helix (HTH) motif. FlhD does not bind DNA by itself, so it may be that the DNA-binding HTH motif becomes rigidly defined only when FlhD forms a complex with some other protein, such as FlhC. If this were true, it might explain how FlhD exhibits plasticity in its DNA-binding specificity, as each partner protein with which it forms a complex could allosterically affect the binding specificity of its HTH motif. A disulphide bridge is seen between the unique cysteine residues (Cys-65) of FlhD native homodimers. Alanine substitution at Cys-65 does not affect FlhD transcription activator activity, suggesting that the disulphide bond is not necessary for either dimer stability or this function of FlhD. Electrostatic potential analysis indicates that dimeric FlhD has a negatively charged surface. |
| | | | |
| - | ==About this Structure==
| + | Crystal structure of the global regulator FlhD from Escherichia coli at 1.8 A resolution.,Campos A, Zhang RG, Alkire RW, Matsumura P, Westbrook EM Mol Microbiol. 2001 Feb;39(3):567-80. PMID:11169099<ref>PMID:11169099</ref> |
| - | 1G8E is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli]. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1G8E OCA].
| + | |
| | | | |
| - | ==Reference==
| + | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> |
| - | Crystal structure of the global regulator FlhD from Escherichia coli at 1.8 A resolution., Campos A, Zhang RG, Alkire RW, Matsumura P, Westbrook EM, Mol Microbiol. 2001 Feb;39(3):567-80. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=11169099 11169099]
| + | </div> |
| - | [[Category: Escherichia coli]]
| + | <div class="pdbe-citations 1g8e" style="background-color:#fffaf0;"></div> |
| - | [[Category: Single protein]]
| + | |
| - | [[Category: Alkire, R.W.]]
| + | |
| - | [[Category: Campos, A.]]
| + | |
| - | [[Category: Matsumura, P.]]
| + | |
| - | [[Category: Westbrook, E.M.]]
| + | |
| - | [[Category: Zhang, R.G.]]
| + | |
| - | [[Category: dna binding protein]]
| + | |
| - | [[Category: genetic regulator]]
| + | |
| | | | |
| - | ''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Tue Nov 20 15:48:39 2007''
| + | ==See Also== |
| | + | *[[Transcriptional activator 3D structures|Transcriptional activator 3D structures]] |
| | + | == References == |
| | + | <references/> |
| | + | __TOC__ |
| | + | </StructureSection> |
| | + | [[Category: Escherichia coli]] |
| | + | [[Category: Large Structures]] |
| | + | [[Category: Alkire RW]] |
| | + | [[Category: Campos A]] |
| | + | [[Category: Matsumura P]] |
| | + | [[Category: Westbrook EM]] |
| | + | [[Category: Zhang RG]] |
| Structural highlights
Function
FLHD_ECOLI Functions in complex with FlhC as a master transcriptional regulator that regulates transcription of several flagellar and non-flagellar operons by binding to their promoter region. Activates expression of class 2 flagellar genes, including fliA, which is a flagellum-specific sigma factor that turns on the class 3 genes. Also regulates genes whose products function in a variety of physiological pathways.[1] [2] [3] [4]
Evolutionary Conservation
Check, as determined by ConSurfDB. You may read the explanation of the method and the full data available from ConSurf.
Publication Abstract from PubMed
FlhD is a 13.3 kDa transcriptional activator protein of flagellar genes and a global regulator. FlhD activates the transcription of class II operons in the flagellar regulon when complexed with a second protein FlhC (21.5 kDa). FlhD also regulates other expression systems in Escherichia coli. We are seeking to understand this plasticity of FlhD's DNA-binding specificity and, to this end, we have determined the crystal structure of the isolated FlhD protein. The structure was solved by substituting seleno-methionine for natural sulphur-methionine in FlhD, crystallizing the protein and determining the structure factor phases by the method of multiple-energy anomalous dispersion (MAD). The FlhD protein is dimeric. The dimer is tightly coupled, with an intimate contact surface, implying that the dimer does not easily dissociate. The FlhD monomer is predominantly alpha-helical. The C-termini of both FlhD monomers (residues 83-116) are completely disrupted by crystal packing, implying that this region of FlhD is highly flexible. However, part of the C-terminus structure in chain A (residues 83-98) was modelled using a native FlhD crystal. What is seen in chain A suggests a classic DNA-binding, helix-turn-helix (HTH) motif. FlhD does not bind DNA by itself, so it may be that the DNA-binding HTH motif becomes rigidly defined only when FlhD forms a complex with some other protein, such as FlhC. If this were true, it might explain how FlhD exhibits plasticity in its DNA-binding specificity, as each partner protein with which it forms a complex could allosterically affect the binding specificity of its HTH motif. A disulphide bridge is seen between the unique cysteine residues (Cys-65) of FlhD native homodimers. Alanine substitution at Cys-65 does not affect FlhD transcription activator activity, suggesting that the disulphide bond is not necessary for either dimer stability or this function of FlhD. Electrostatic potential analysis indicates that dimeric FlhD has a negatively charged surface.
Crystal structure of the global regulator FlhD from Escherichia coli at 1.8 A resolution.,Campos A, Zhang RG, Alkire RW, Matsumura P, Westbrook EM Mol Microbiol. 2001 Feb;39(3):567-80. PMID:11169099[5]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Liu X, Matsumura P. The FlhD/FlhC complex, a transcriptional activator of the Escherichia coli flagellar class II operons. J Bacteriol. 1994 Dec;176(23):7345-51. PMID:7961507
- ↑ Campos A, Matsumura P. Extensive alanine scanning reveals protein-protein and protein-DNA interaction surfaces in the global regulator FlhD from Escherichia coli. Mol Microbiol. 2001 Feb;39(3):581-94. PMID:11169100
- ↑ Stafford GP, Ogi T, Hughes C. Binding and transcriptional activation of non-flagellar genes by the Escherichia coli flagellar master regulator FlhD2C2. Microbiology. 2005 Jun;151(Pt 6):1779-88. PMID:15941987 doi:10.1099/mic.0.27879-0
- ↑ Pesavento C, Becker G, Sommerfeldt N, Possling A, Tschowri N, Mehlis A, Hengge R. Inverse regulatory coordination of motility and curli-mediated adhesion in Escherichia coli. Genes Dev. 2008 Sep 1;22(17):2434-46. doi: 10.1101/gad.475808. PMID:18765794 doi:10.1101/gad.475808
- ↑ Campos A, Zhang RG, Alkire RW, Matsumura P, Westbrook EM. Crystal structure of the global regulator FlhD from Escherichia coli at 1.8 A resolution. Mol Microbiol. 2001 Feb;39(3):567-80. PMID:11169099
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