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| | <SX load='3j7h' size='340' side='right' viewer='molstar' caption='[[3j7h]], [[Resolution|resolution]] 3.20Å' scene=''> | | <SX load='3j7h' size='340' side='right' viewer='molstar' caption='[[3j7h]], [[Resolution|resolution]] 3.20Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[3j7h]] is a 4 chain structure with sequence from [http://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=3J7H OCA]. For a <b>guided tour on the structure components</b> use [http://proteopedia.org/fgij/fg.htm?mol=3J7H FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[3j7h]] is a 4 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=3J7H OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3J7H FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene></td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">Electron Microscopy, [[Resolution|Resolution]] 3.2Å</td></tr> |
| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Beta-galactosidase Beta-galactosidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.23 3.2.1.23] </span></td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene></td></tr> |
| - | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://proteopedia.org/fgij/fg.htm?mol=3j7h FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3j7h OCA], [http://pdbe.org/3j7h PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=3j7h RCSB], [http://www.ebi.ac.uk/pdbsum/3j7h PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=3j7h 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=3j7h FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3j7h OCA], [https://pdbe.org/3j7h PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3j7h RCSB], [https://www.ebi.ac.uk/pdbsum/3j7h PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3j7h ProSAT]</span></td></tr> |
| | </table> | | </table> |
| - | <div style="background-color:#fffaf0;">
| + | == Function == |
| - | == Publication Abstract from PubMed == | + | [https://www.uniprot.org/uniprot/BGAL_ECOLI BGAL_ECOLI] |
| - | We report the solution structure of Escherichia coli beta-galactosidase ( approximately 465 kDa), solved at approximately 3.2-A resolution by using single-particle cryo-electron microscopy (cryo-EM). Densities for most side chains, including those of residues in the active site, and a catalytic Mg2+ ion can be discerned in the map obtained by cryo-EM. The atomic model derived from our cryo-EM analysis closely matches the 1.7-A crystal structure with a global rmsd of approximately 0.66 A. There are significant local differences throughout the protein, with clear evidence for conformational changes resulting from contact zones in the crystal lattice. Inspection of the map reveals that although densities for residues with positively charged and neutral side chains are well resolved, systematically weaker densities are observed for residues with negatively charged side chains. We show that the weaker densities for negatively charged residues arise from their greater sensitivity to radiation damage from electron irradiation as determined by comparison of density maps obtained by using electron doses ranging from 10 to 30 e-/A2. In summary, we establish that it is feasible to use cryo-EM to determine near-atomic resolution structures of protein complexes (<500 kDa) with low symmetry, and that the residue-specific radiation damage that occurs with increasing electron dose can be monitored by using dose fractionation tools available with direct electron detector technology.
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| - | | + | |
| - | Structure of beta-galactosidase at 3.2-A resolution obtained by cryo-electron microscopy.,Bartesaghi A, Matthies D, Banerjee S, Merk A, Subramaniam S Proc Natl Acad Sci U S A. 2014 Jul 28. pii: 201402809. PMID:25071206<ref>PMID:25071206</ref>
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| - | | + | |
| - | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
| + | |
| - | </div>
| + | |
| - | <div class="pdbe-citations 3j7h" style="background-color:#fffaf0;"></div>
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| | | | |
| | ==See Also== | | ==See Also== |
| | *[[Galactosidase 3D structures|Galactosidase 3D structures]] | | *[[Galactosidase 3D structures|Galactosidase 3D structures]] |
| - | == References == | |
| - | <references/> | |
| | __TOC__ | | __TOC__ |
| | </SX> | | </SX> |
| - | [[Category: Beta-galactosidase]]
| + | [[Category: Escherichia coli K-12]] |
| - | [[Category: Escherichia coli k-12]] | + | |
| | [[Category: Large Structures]] | | [[Category: Large Structures]] |
| - | [[Category: Banerjee, S]] | + | [[Category: Banerjee S]] |
| - | [[Category: Bartesaghi, A]] | + | [[Category: Bartesaghi A]] |
| - | [[Category: Matthies, D]] | + | [[Category: Matthies D]] |
| - | [[Category: Merk, A]] | + | [[Category: Merk A]] |
| - | [[Category: Subramaniam, S]] | + | [[Category: Subramaniam S]] |
| - | [[Category: 3d reconstruction]]
| + | |
| - | [[Category: Atomic resolution cryo-electron microscopy]]
| + | |
| - | [[Category: Direct electron detector]]
| + | |
| - | [[Category: Homo-tetramer]]
| + | |
| - | [[Category: Hydrolase]]
| + | |
| - | [[Category: Hydrolase enzyme]]
| + | |
| - | [[Category: Protein complex]]
| + | |
| - | [[Category: Single-particle cryo-em]]
| + | |
