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3w0p

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{{STRUCTURE_3w0p| PDB=3w0p | SCENE= }}
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==Crystal structure of a thermostable mutant of aminoglycoside phosphotransferase APH(4)-Ia (D198A), ternary complex with ADP and hygromycin B==
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===Crystal structure of a thermostable mutant of aminoglycoside phosphotransferase APH(4)-Ia (D198A), ternary complex with ADP and hygromycin B===
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<StructureSection load='3w0p' size='340' side='right' caption='[[3w0p]], [[Resolution|resolution]] 2.00&Aring;' scene=''>
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{{ABSTRACT_PUBMED_23747390}}
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== Structural highlights ==
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<table><tr><td colspan='2'>[[3w0p]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3W0P OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=3W0P FirstGlance]. <br>
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</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=ADP:ADENOSINE-5-DIPHOSPHATE'>ADP</scene>, <scene name='pdbligand=HY0:HYGROMYCIN+B+VARIANT'>HY0</scene></td></tr>
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<tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[3w0m|3w0m]], [[3w0n|3w0n]], [[3w0o|3w0o]], [[3w0q|3w0q]], [[3w0r|3w0r]], [[3w0s|3w0s]]</td></tr>
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<tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">hph ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=562 Escherichia coli])</td></tr>
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<tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Hygromycin_B_4-O-kinase Hygromycin B 4-O-kinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.1.163 2.7.1.163] </span></td></tr>
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<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=3w0p FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3w0p OCA], [http://www.rcsb.org/pdb/explore.do?structureId=3w0p RCSB], [http://www.ebi.ac.uk/pdbsum/3w0p PDBsum]</span></td></tr>
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</table>
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<div style="background-color:#fffaf0;">
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== Publication Abstract from PubMed ==
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Aminoglycoside 4-phosphotransferase-Ia (APH(4)-Ia)/Hygromycin B phosphotransferase (Hph) inactivates the aminoglycoside antibiotic hygromycin B (hygB) via phosphorylation. The crystal structure of the binary complex of APH(4)-Ia with hygB was recently reported. To characterize substrate recognition by the enzyme, we determined the crystal structure of the ternary complex of non-hydrolyzable ATP analog AMP-PNP and hygB with wild-type, thermostable Hph mutant Hph5, and apo-mutant enzyme forms. The comparison between the ternary complex and apo structures revealed that Hph undergoes domain movement upon binding of AMP-PNP and hygB. This was about half amount of the case of APH(9)-Ia. We also determined the crystal structures of mutants in which the conserved, catalytically important residues Asp198 and Asn203, and the non-conserved Asn202, were converted to Ala, revealing the importance of Asn202 for catalysis. Hph5 contains five amino acid substitutions that alter its thermostability by 16 degrees C; its structure revealed that 4/5 mutations in Hph5 are located in the hydrophobic core and appear to increase thermostability by strengthening hydrophobic interactions.
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==Function==
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Crystal structures of the ternary complex of APH(4)-Ia/Hph with hygromycin B and an ATP analog using a thermostable mutant.,Iino D, Takakura Y, Fukano K, Sasaki Y, Hoshino T, Ohsawa K, Nakamura A, Yajima S J Struct Biol. 2013 Jul;183(1):76-85. doi: 10.1016/j.jsb.2013.05.023. Epub 2013, Jun 7. PMID:23747390<ref>PMID:23747390</ref>
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[[http://www.uniprot.org/uniprot/KHYB_ECOLX KHYB_ECOLX]] The aminoglycoside phosphotransferases achieve inactivation of their antibiotic substrates by phosphorylation. Only phosphorylates hygromycin and closely related compounds such as demethyl analogs and destomycin.
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==About this Structure==
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From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
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[[3w0p]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3W0P OCA].
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</div>
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== References ==
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==Reference==
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<references/>
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<ref group="xtra">PMID:023747390</ref><references group="xtra"/><references/>
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__TOC__
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</StructureSection>
[[Category: Escherichia coli]]
[[Category: Escherichia coli]]
[[Category: Hygromycin B 4-O-kinase]]
[[Category: Hygromycin B 4-O-kinase]]
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[[Category: Fukano, K.]]
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[[Category: Fukano, K]]
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[[Category: Hoshino, T.]]
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[[Category: Hoshino, T]]
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[[Category: Iino, D.]]
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[[Category: Iino, D]]
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[[Category: Nakamura, A.]]
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[[Category: Nakamura, A]]
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[[Category: Ohsawa, K.]]
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[[Category: Ohsawa, K]]
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[[Category: Sasaki, Y.]]
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[[Category: Sasaki, Y]]
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[[Category: Takakura, Y.]]
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[[Category: Takakura, Y]]
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[[Category: Yajima, S.]]
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[[Category: Yajima, S]]
[[Category: Phosphotransferase]]
[[Category: Phosphotransferase]]
[[Category: Transferase-antibiotic complex]]
[[Category: Transferase-antibiotic complex]]

Revision as of 07:35, 21 December 2014

Crystal structure of a thermostable mutant of aminoglycoside phosphotransferase APH(4)-Ia (D198A), ternary complex with ADP and hygromycin B

3w0p, resolution 2.00Å

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