1fxh: Difference between revisions

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New page: left|200px<br /><applet load="1fxh" size="450" color="white" frame="true" align="right" spinBox="true" caption="1fxh, resolution 1.97Å" /> '''MUTANT OF PENICILLIN...
 
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[[Image:1fxh.jpg|left|200px]]<br /><applet load="1fxh" size="450" color="white" frame="true" align="right" spinBox="true"
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'''MUTANT OF PENICILLIN ACYLASE IMPAIRED IN CATALYSIS WITH PHENYLACETIC ACID IN THE ACTIVE SITE'''<br />


==Overview==
==MUTANT OF PENICILLIN ACYLASE IMPAIRED IN CATALYSIS WITH PHENYLACETIC ACID IN THE ACTIVE SITE==
The binding of penicillin to penicillin acylase was studied by X-ray, crystallography. The structure of the enzyme-substrate complex was, determined after soaking crystals of an inactive betaN241A penicillin, acylase mutant with penicillin G. Binding of the substrate induces a, conformational change, in which the side chains of alphaF146 and alphaR145, move away from the active site, which allows the enzyme to accommodate, penicillin G. In the resulting structure, the beta-lactam binding site is, formed by the side chains of alphaF146 and betaF71, which have van der, Waals interactions with the thiazolidine ring of penicillin G and the side, chain of alphaR145 that is connected to the carboxylate group of the, ligand by means of hydrogen bonding via two water molecules. The backbone, oxygen of betaQ23 forms a hydrogen bond with the carbonyl oxygen of the, phenylacetic acid moiety through a bridging water molecule. Kinetic, studies revealed that the site-directed mutants alphaF146Y, alphaF146A and, alphaF146L all show significant changes in their interaction with the, beta-lactam substrates as compared with the wild type. The alphaF146Y, mutant had the same affinity for 6-aminopenicillanic acid as the wild-type, enzyme, but was not able to synthesize penicillin G from phenylacetamide, and 6-aminopenicillanic acid. The alphaF146L and alphaF146A enzymes had a, 3-5-fold decreased affinity for 6-aminopenicillanic acid, but synthesized, penicillin G more efficiently than the wild type. The combined results of, the structural and kinetic studies show the importance of alphaF146 in the, beta-lactam binding site and provide leads for engineering mutants with, improved synthetic properties.
<StructureSection load='1fxh' size='340' side='right'caption='[[1fxh]], [[Resolution|resolution]] 1.97&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[1fxh]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1FXH OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1FXH FirstGlance]. <br>
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.97&#8491;</td></tr>
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=PAC:2-PHENYLACETIC+ACID'>PAC</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=1fxh FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1fxh OCA], [https://pdbe.org/1fxh PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1fxh RCSB], [https://www.ebi.ac.uk/pdbsum/1fxh PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1fxh ProSAT]</span></td></tr>
</table>
== Function ==
[https://www.uniprot.org/uniprot/PAC_ECOLX PAC_ECOLX]
== Evolutionary Conservation ==
[[Image:Consurf_key_small.gif|200px|right]]
Check<jmol>
  <jmolCheckbox>
    <scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/fx/1fxh_consurf.spt"</scriptWhenChecked>
    <scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview01.spt</scriptWhenUnchecked>
    <text>to colour the structure by Evolutionary Conservation</text>
  </jmolCheckbox>
</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=1fxh ConSurf].
<div style="clear:both"></div>


==About this Structure==
==See Also==
1FXH is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli] with CA and PAC as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Penicillin_amidase Penicillin amidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.5.1.11 3.5.1.11] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1FXH OCA].
*[[Penicillin acylase|Penicillin acylase]]
 
__TOC__
==Reference==
</StructureSection>
Characterization of the beta-lactam binding site of penicillin acylase of Escherichia coli by structural and site-directed mutagenesis studies., Alkema WB, Hensgens CM, Kroezinga EH, de Vries E, Floris R, van der Laan JM, Dijkstra BW, Janssen DB, Protein Eng. 2000 Dec;13(12):857-63. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=11239085 11239085]
[[Category: Escherichia coli]]
[[Category: Escherichia coli]]
[[Category: Penicillin amidase]]
[[Category: Large Structures]]
[[Category: Protein complex]]
[[Category: Alkema WB]]
[[Category: Alkema, W.B.]]
[[Category: Dijkstra BW]]
[[Category: Dijkstra, B.W.]]
[[Category: Floris R]]
[[Category: Floris, R.]]
[[Category: Hensgens CM]]
[[Category: Hensgens, C.M.]]
[[Category: Janssen DB]]
[[Category: Janssen, D.B.]]
[[Category: Kroezinga EH]]
[[Category: Kroezinga, E.H.]]
[[Category: De Vries E]]
[[Category: Laan, J.M.van.der.]]
[[Category: Van der Laan JM]]
[[Category: Vries, E.de.]]
[[Category: CA]]
[[Category: PAC]]
[[Category: ntn-hydrolase fold]]
 
''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Tue Nov 20 15:26:08 2007''

Latest revision as of 10:20, 7 February 2024

MUTANT OF PENICILLIN ACYLASE IMPAIRED IN CATALYSIS WITH PHENYLACETIC ACID IN THE ACTIVE SITEMUTANT OF PENICILLIN ACYLASE IMPAIRED IN CATALYSIS WITH PHENYLACETIC ACID IN THE ACTIVE SITE

Structural highlights

1fxh is a 2 chain structure with sequence from Escherichia coli. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:X-ray diffraction, Resolution 1.97Å
Ligands:,
Resources:FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

PAC_ECOLX

Evolutionary Conservation

Check, as determined by ConSurfDB. You may read the explanation of the method and the full data available from ConSurf.

See Also

1fxh, resolution 1.97Å

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