1rgl

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(New page: 200px<br /><applet load="1rgl" size="450" color="white" frame="true" align="right" spinBox="true" caption="1rgl, resolution 2.0&Aring;" /> '''RNASE T1 MUTANT GLU46...)
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[[Image:1rgl.gif|left|200px]]<br /><applet load="1rgl" size="450" color="white" frame="true" align="right" spinBox="true"
 
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caption="1rgl, resolution 2.0&Aring;" />
 
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'''RNASE T1 MUTANT GLU46GLN BINDS THE INHIBITORS 2'GMP AND 2'AMP AT THE 3' SUBSITE'''<br />
 
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==Overview==
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==RNASE T1 MUTANT GLU46GLN BINDS THE INHIBITORS 2'GMP AND 2'AMP AT THE 3' SUBSITE==
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On the basis of molecular dynamics and free-energy perturbation, approaches, the Glu46Gln (E46Q) mutation in the guanine-specific, ribonuclease T1 (RNase T1) was predicted to render the enzyme specific for, adenine. The E46Q mutant was genetically engineered and characterized, biochemically and crystallographically by investigating the structures of, its two complexes with 2'AMP and 2'GMP. The ribonuclease E46Q mutant is, nearly inactive towards dinucleoside phosphate substrates but shows 17%, residual activity towards RNA. It binds 2'AMP and 2'GMP equally well with, dissociation constants of 49 microM and 37 microM, in contrast to the, wild-type enzyme, which strongly discriminates between these two, nucleotides, yielding dissociation constants of 36 microM and 0.6 microM., These data suggest that the E46Q mutant binds the nucleotides not to the, specific recognition site but to the subsite at His92. This was confirmed, by the crystal structures, which also showed that the Gln46 amide is, hydrogen bonded to the Phe100 N and O atoms, and tightly anchored in this, position. This interaction may either have locked the guanine recognition, site so that 2'AMP and 2'GMP are unable to insert, or the contribution to, guanine recognition of Glu46 is so important that the E46Q mutant is, unable to function in recognition of either guanine and adenine.
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<StructureSection load='1rgl' size='340' side='right'caption='[[1rgl]], [[Resolution|resolution]] 2.00&Aring;' scene=''>
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== Structural highlights ==
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<table><tr><td colspan='2'>[[1rgl]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Aspergillus_oryzae Aspergillus oryzae]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1RGL OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1RGL FirstGlance]. <br>
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</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2&#8491;</td></tr>
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<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=2GP:GUANOSINE-2-MONOPHOSPHATE'>2GP</scene>, <scene name='pdbligand=CA:CALCIUM+ION'>CA</scene></td></tr>
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<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=1rgl FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1rgl OCA], [https://pdbe.org/1rgl PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1rgl RCSB], [https://www.ebi.ac.uk/pdbsum/1rgl PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1rgl ProSAT]</span></td></tr>
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</table>
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== Function ==
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[https://www.uniprot.org/uniprot/RNT1_ASPOR RNT1_ASPOR]
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== Evolutionary Conservation ==
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[[Image:Consurf_key_small.gif|200px|right]]
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Check<jmol>
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<jmolCheckbox>
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<scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/rg/1rgl_consurf.spt"</scriptWhenChecked>
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<scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview03.spt</scriptWhenUnchecked>
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<text>to colour the structure by Evolutionary Conservation</text>
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</jmolCheckbox>
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</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=1rgl ConSurf].
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<div style="clear:both"></div>
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<div style="background-color:#fffaf0;">
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== Publication Abstract from PubMed ==
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On the basis of molecular dynamics and free-energy perturbation approaches, the Glu46Gln (E46Q) mutation in the guanine-specific ribonuclease T1 (RNase T1) was predicted to render the enzyme specific for adenine. The E46Q mutant was genetically engineered and characterized biochemically and crystallographically by investigating the structures of its two complexes with 2'AMP and 2'GMP. The ribonuclease E46Q mutant is nearly inactive towards dinucleoside phosphate substrates but shows 17% residual activity towards RNA. It binds 2'AMP and 2'GMP equally well with dissociation constants of 49 microM and 37 microM, in contrast to the wild-type enzyme, which strongly discriminates between these two nucleotides, yielding dissociation constants of 36 microM and 0.6 microM. These data suggest that the E46Q mutant binds the nucleotides not to the specific recognition site but to the subsite at His92. This was confirmed by the crystal structures, which also showed that the Gln46 amide is hydrogen bonded to the Phe100 N and O atoms, and tightly anchored in this position. This interaction may either have locked the guanine recognition site so that 2'AMP and 2'GMP are unable to insert, or the contribution to guanine recognition of Glu46 is so important that the E46Q mutant is unable to function in recognition of either guanine and adenine.
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==About this Structure==
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RNase T1 mutant Glu46Gln binds the inhibitors 2'GMP and 2'AMP at the 3' subsite.,Granzin J, Puras-Lutzke R, Landt O, Grunert HP, Heinemann U, Saenger W, Hahn U J Mol Biol. 1992 May 20;225(2):533-42. PMID:1350642<ref>PMID:1350642</ref>
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1RGL is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Aspergillus_oryzae Aspergillus oryzae] with CA and 2GP as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Ribonuclease_T(1) Ribonuclease T(1)], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.27.3 3.1.27.3] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1RGL OCA].
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==Reference==
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From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
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RNase T1 mutant Glu46Gln binds the inhibitors 2'GMP and 2'AMP at the 3' subsite., Granzin J, Puras-Lutzke R, Landt O, Grunert HP, Heinemann U, Saenger W, Hahn U, J Mol Biol. 1992 May 20;225(2):533-42. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=1350642 1350642]
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</div>
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[[Category: Aspergillus oryzae]]
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<div class="pdbe-citations 1rgl" style="background-color:#fffaf0;"></div>
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[[Category: Ribonuclease T(1)]]
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[[Category: Single protein]]
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[[Category: Granzin, J.]]
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[[Category: Grunert, H.P.]]
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[[Category: Hahn, U.]]
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[[Category: Heinemann, U.]]
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[[Category: Landt, O.]]
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[[Category: Puras-Lutzke, R.]]
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[[Category: Saenger, W.]]
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[[Category: 2GP]]
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[[Category: CA]]
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[[Category: hydrolase(endoribonuclease)]]
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''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Wed Nov 21 01:34:56 2007''
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==See Also==
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*[[Ribonuclease 3D structures|Ribonuclease 3D structures]]
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== References ==
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<references/>
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__TOC__
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</StructureSection>
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[[Category: Aspergillus oryzae]]
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[[Category: Large Structures]]
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[[Category: Granzin J]]
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[[Category: Grunert H-P]]
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[[Category: Hahn U]]
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[[Category: Heinemann U]]
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[[Category: Landt O]]
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[[Category: Puras-Lutzke R]]
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[[Category: Saenger W]]

Current revision

RNASE T1 MUTANT GLU46GLN BINDS THE INHIBITORS 2'GMP AND 2'AMP AT THE 3' SUBSITE

PDB ID 1rgl

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