3w0p: Difference between revisions
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==Crystal structure of a thermostable mutant of aminoglycoside phosphotransferase APH(4)-Ia (D198A), ternary complex with ADP and hygromycin B== | |||
<StructureSection load='3w0p' size='340' side='right' caption='[[3w0p]], [[Resolution|resolution]] 2.00Å' scene=''> | |||
== Structural highlights == | |||
<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> | |||
</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> | |||
<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> | |||
<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> | |||
<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> | |||
<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> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
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. | |||
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> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
== References == | |||
== | <references/> | ||
__TOC__ | |||
</StructureSection> | |||
[[Category: Escherichia coli]] | [[Category: Escherichia coli]] | ||
[[Category: Hygromycin B 4-O-kinase]] | [[Category: Hygromycin B 4-O-kinase]] | ||
[[Category: Fukano, K | [[Category: Fukano, K]] | ||
[[Category: Hoshino, T | [[Category: Hoshino, T]] | ||
[[Category: Iino, D | [[Category: Iino, D]] | ||
[[Category: Nakamura, A | [[Category: Nakamura, A]] | ||
[[Category: Ohsawa, K | [[Category: Ohsawa, K]] | ||
[[Category: Sasaki, Y | [[Category: Sasaki, Y]] | ||
[[Category: Takakura, Y | [[Category: Takakura, Y]] | ||
[[Category: Yajima, S | [[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
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