5cfu: Difference between revisions
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==Crystal Structure of ANT(2")-Ia in complex with adenylyl-2"-tobramycin== | |||
<StructureSection load='5cfu' size='340' side='right'caption='[[5cfu]], [[Resolution|resolution]] 1.82Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[5cfu]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Pseudomonas_aeruginosa Pseudomonas aeruginosa]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5CFU OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5CFU FirstGlance]. <br> | |||
</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=51H:ADENYLYL-2+-TOBRAMYCIN'>51H</scene>, <scene name='pdbligand=BU1:1,4-BUTANEDIOL'>BU1</scene>, <scene name='pdbligand=MN:MANGANESE+(II)+ION'>MN</scene>, <scene name='pdbligand=POP:PYROPHOSPHATE+2-'>POP</scene></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=5cfu FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5cfu OCA], [https://pdbe.org/5cfu PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5cfu RCSB], [https://www.ebi.ac.uk/pdbsum/5cfu PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5cfu ProSAT]</span></td></tr> | |||
</table> | |||
== Function == | |||
[https://www.uniprot.org/uniprot/Q6X3H6_PSEAI Q6X3H6_PSEAI] | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Aminoglycoside antibiotics have lost much of their effectiveness due to widespread resistance, primarily via covalent modification. One of the most ubiquitous enzymes responsible for aminoglycoside resistance is aminoglycoside O-nucleotidyltransferase(2''), which catalyzes a nucleotidylation reaction. Due to its clinical importance, much research has focused on dissecting the mechanism of action, some of it dating back more than 30 years. Here, we present structural data for catalytically informative states of the enzyme, i.e., ANT(2'') in complex with adenosine monophosphate (AMP) and tobramycin (inactive-intermediate state) and in complex with adenylyl-2''-tobramycin, pyrophosphate, and Mn(2+)(product-bound state). These two structures in conjunction with our previously reported structure of ANT(2'')'s substrate-bound complex capture clinical states along ANT(2'')'s reaction coordinate. Additionally, isothermal titration calorimetry (ITC)-based studies are presented that assess the order of substrate binding and product release. Combined, these results outline a kinetic mechanism for ANT(2'') that contradicts what has been previously reported. Specifically, we show that the release of adenylated aminoglycoside precedes pyrophosphate. Furthermore, the ternary complex structures provide additional details on the catalytic mechanism, which reveals extensive similarities to the evolutionarily related DNA polymerase-beta superfamily. | |||
Revisiting the Catalytic Cycle and Kinetic Mechanism of Aminoglycoside O-Nucleotidyltransferase(2''): A Structural and Kinetic Study.,Bassenden AV, Park J, Rodionov D, Berghuis AM ACS Chem Biol. 2020 Mar 6. doi: 10.1021/acschembio.9b00904. PMID:32100995<ref>PMID:32100995</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[Category: | </div> | ||
[[Category: Bassenden | <div class="pdbe-citations 5cfu" style="background-color:#fffaf0;"></div> | ||
[[Category: | == References == | ||
[[Category: | <references/> | ||
__TOC__ | |||
</StructureSection> | |||
[[Category: Large Structures]] | |||
[[Category: Pseudomonas aeruginosa]] | |||
[[Category: Bassenden AV]] | |||
[[Category: Berghuis AM]] | |||
[[Category: Rodionov D]] | |||
Latest revision as of 15:03, 14 June 2023
Crystal Structure of ANT(2")-Ia in complex with adenylyl-2"-tobramycinCrystal Structure of ANT(2")-Ia in complex with adenylyl-2"-tobramycin
Structural highlights
FunctionPublication Abstract from PubMedAminoglycoside antibiotics have lost much of their effectiveness due to widespread resistance, primarily via covalent modification. One of the most ubiquitous enzymes responsible for aminoglycoside resistance is aminoglycoside O-nucleotidyltransferase(2), which catalyzes a nucleotidylation reaction. Due to its clinical importance, much research has focused on dissecting the mechanism of action, some of it dating back more than 30 years. Here, we present structural data for catalytically informative states of the enzyme, i.e., ANT(2) in complex with adenosine monophosphate (AMP) and tobramycin (inactive-intermediate state) and in complex with adenylyl-2-tobramycin, pyrophosphate, and Mn(2+)(product-bound state). These two structures in conjunction with our previously reported structure of ANT(2)'s substrate-bound complex capture clinical states along ANT(2)'s reaction coordinate. Additionally, isothermal titration calorimetry (ITC)-based studies are presented that assess the order of substrate binding and product release. Combined, these results outline a kinetic mechanism for ANT(2) that contradicts what has been previously reported. Specifically, we show that the release of adenylated aminoglycoside precedes pyrophosphate. Furthermore, the ternary complex structures provide additional details on the catalytic mechanism, which reveals extensive similarities to the evolutionarily related DNA polymerase-beta superfamily. Revisiting the Catalytic Cycle and Kinetic Mechanism of Aminoglycoside O-Nucleotidyltransferase(2): A Structural and Kinetic Study.,Bassenden AV, Park J, Rodionov D, Berghuis AM ACS Chem Biol. 2020 Mar 6. doi: 10.1021/acschembio.9b00904. PMID:32100995[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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