1gk0

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[[Image:1gk0.gif|left|200px]]<br />
 
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<applet load="1gk0" size="450" color="white" frame="true" align="right" spinBox="true"
 
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caption="1gk0, resolution 2.50&Aring;" />
 
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'''STRUCTURE-BASED PREDICTION OF MODIFICATIONS IN GLUTARYLAMIDASE TO ALLOW SINGLE-STEP ENZYMATIC PRODUCTION OF 7-AMINOCEPHALOSPORANIC ACID FROM CEPHALOSPORIN C'''<br />
 
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==Overview==
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==Structure-based prediction of modifications in glutarylamidase to allow single-step enzymatic production of 7-aminocephalosporanic acid from cephalosporin C==
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Glutarylamidase is an important enzyme employed in the commercial, production of 7-aminocephalosporanic acid, a starting compound in the, synthesis of cephalosporin antibiotics. 7-aminocephalosporanic acid is, obtained from cephalosporin C, a natural antibiotic, either chemically or, by a two-step enzymatic process utilizing the enzymes D-amino acid oxidase, and glutarylamidase. We have investigated possibilities for redesigning, glutarylamidase for the production of 7-aminocephalosporanic acid from, cephalosporin C in a single enzymatic step. These studies are based on the, structures of glutarylamidase, which we have solved with bound phosphate, and ethylene glycol to 2.5 A resolution and with bound glycerol to 2.4 A., The phosphate binds near the catalytic serine in a way that mimics the, hemiacetal that develops during catalysis, while the glycerol occupies the, side-chain binding pocket. Our structures show that the enzyme is not only, structurally similar to penicillin G acylase but also employs essentially, the same mechanism in which the alpha-amino group of the catalytic serine, acts as a base. A subtle difference is the presence of two catalytic, dyads, His B23/Glu B455 and His B23/Ser B1, that are not seen in, penicillin G acylase. In contrast to classical serine proteases, the, central histidine of these dyads interacts indirectly with the O(gamma), through a hydrogen bond relay network involving the alpha-amino group of, the serine and a bound water molecule. A plausible model of the, enzyme-substrate complex is proposed that leads to the prediction of, mutants of glutarylamidase that should enable the enzyme to deacylate, cephalosporin C into 7-aminocephalosporanic acid.
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<StructureSection load='1gk0' size='340' side='right'caption='[[1gk0]], [[Resolution|resolution]] 2.50&Aring;' scene=''>
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== Structural highlights ==
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<table><tr><td colspan='2'>[[1gk0]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Pseudomonas_sp. Pseudomonas sp.]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1GK0 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1GK0 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]] 2.5&#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=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=MSE:SELENOMETHIONINE'>MSE</scene>, <scene name='pdbligand=PO4:PHOSPHATE+ION'>PO4</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=1gk0 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1gk0 OCA], [https://pdbe.org/1gk0 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1gk0 RCSB], [https://www.ebi.ac.uk/pdbsum/1gk0 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1gk0 ProSAT]</span></td></tr>
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</table>
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== Function ==
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[https://www.uniprot.org/uniprot/G7AC_PSEU7 G7AC_PSEU7] Catalyzes the deacylation of 7 beta-(4-carboxybutanamido)cephalosporanic acid (glutaryl-7-aminocephalosporanic acid or GL-7-ACA) to 7-aminocephalosporanic acid (7-ACA).<ref>PMID:2993240</ref>
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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/gk/1gk0_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=1gk0 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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Glutarylamidase is an important enzyme employed in the commercial production of 7-aminocephalosporanic acid, a starting compound in the synthesis of cephalosporin antibiotics. 7-aminocephalosporanic acid is obtained from cephalosporin C, a natural antibiotic, either chemically or by a two-step enzymatic process utilizing the enzymes D-amino acid oxidase and glutarylamidase. We have investigated possibilities for redesigning glutarylamidase for the production of 7-aminocephalosporanic acid from cephalosporin C in a single enzymatic step. These studies are based on the structures of glutarylamidase, which we have solved with bound phosphate and ethylene glycol to 2.5 A resolution and with bound glycerol to 2.4 A. The phosphate binds near the catalytic serine in a way that mimics the hemiacetal that develops during catalysis, while the glycerol occupies the side-chain binding pocket. Our structures show that the enzyme is not only structurally similar to penicillin G acylase but also employs essentially the same mechanism in which the alpha-amino group of the catalytic serine acts as a base. A subtle difference is the presence of two catalytic dyads, His B23/Glu B455 and His B23/Ser B1, that are not seen in penicillin G acylase. In contrast to classical serine proteases, the central histidine of these dyads interacts indirectly with the O(gamma) through a hydrogen bond relay network involving the alpha-amino group of the serine and a bound water molecule. A plausible model of the enzyme-substrate complex is proposed that leads to the prediction of mutants of glutarylamidase that should enable the enzyme to deacylate cephalosporin C into 7-aminocephalosporanic acid.
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==About this Structure==
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Structure-based prediction of modifications in glutarylamidase to allow single-step enzymatic production of 7-aminocephalosporanic acid from cephalosporin C.,Fritz-Wolf K, Koller KP, Lange G, Liesum A, Sauber K, Schreuder H, Aretz W, Kabsch W Protein Sci. 2002 Jan;11(1):92-103. PMID:11742126<ref>PMID:11742126</ref>
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1GK0 is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Pseudomonas_sp. Pseudomonas sp.] with PO4 and EDO 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] Structure known Active Site: POB. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1GK0 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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Structure-based prediction of modifications in glutarylamidase to allow single-step enzymatic production of 7-aminocephalosporanic acid from cephalosporin C., Fritz-Wolf K, Koller KP, Lange G, Liesum A, Sauber K, Schreuder H, Aretz W, Kabsch W, Protein Sci. 2002 Jan;11(1):92-103. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=11742126 11742126]
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</div>
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[[Category: Penicillin amidase]]
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<div class="pdbe-citations 1gk0" style="background-color:#fffaf0;"></div>
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[[Category: Protein complex]]
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[[Category: Pseudomonas sp.]]
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[[Category: Aretz, W.]]
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[[Category: Fritz-Wolf, K.]]
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[[Category: Kabsch, W.]]
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[[Category: Koller, K.P.]]
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[[Category: Lange, G.]]
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[[Category: Liesum, A.]]
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[[Category: Sauber, K.]]
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[[Category: Schreuder, H.]]
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[[Category: EDO]]
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[[Category: PO4]]
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[[Category: catalytic triad]]
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[[Category: cephalosporin acylase]]
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[[Category: cephalosporin c]]
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[[Category: glutaryl acylase]]
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[[Category: ntn-hydrolase]]
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[[Category: x-raz structure]]
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''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Mon Nov 5 12:31:07 2007''
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==See Also==
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*[[Cephalosporin acylase 3D structures|Cephalosporin acylase 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: Large Structures]]
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[[Category: Pseudomonas sp]]
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[[Category: Aretz W]]
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[[Category: Fritz-Wolf K]]
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[[Category: Kabsch W]]
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[[Category: Koller KP]]
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[[Category: Lange G]]
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[[Category: Liesum A]]
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[[Category: Sauber K]]
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[[Category: Schreuder H]]

Current revision

Structure-based prediction of modifications in glutarylamidase to allow single-step enzymatic production of 7-aminocephalosporanic acid from cephalosporin C

PDB ID 1gk0

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