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| | == Structural highlights == | | == Structural highlights == |
| | <table><tr><td colspan='2'>[[7poy]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Aspergillus_nidulans_FGSC_A4 Aspergillus nidulans FGSC A4]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7POY OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7POY FirstGlance]. <br> | | <table><tr><td colspan='2'>[[7poy]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Aspergillus_nidulans_FGSC_A4 Aspergillus nidulans FGSC A4]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7POY OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7POY FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=81T:~{N}-[(3~{R})-2,2,5,5-tetramethyl-1-oxidanyl-pyrrolidin-3-yl]ethanamide'>81T</scene>, <scene name='pdbligand=ACV:L-D-(A-AMINOADIPOYL)-L-CYSTEINYL-D-VALINE'>ACV</scene>, <scene name='pdbligand=FE:FE+(III)+ION'>FE</scene>, <scene name='pdbligand=NO:NITRIC+OXIDE'>NO</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.75Å</td></tr> |
| | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=81T:~{N}-[(3~{R})-2,2,5,5-tetramethyl-1-oxidanyl-pyrrolidin-3-yl]ethanamide'>81T</scene>, <scene name='pdbligand=ACV:L-D-(A-AMINOADIPOYL)-L-CYSTEINYL-D-VALINE'>ACV</scene>, <scene name='pdbligand=FE:FE+(III)+ION'>FE</scene>, <scene name='pdbligand=NO:NITRIC+OXIDE'>NO</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</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=7poy FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7poy OCA], [https://pdbe.org/7poy PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7poy RCSB], [https://www.ebi.ac.uk/pdbsum/7poy PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7poy ProSAT]</span></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=7poy FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7poy OCA], [https://pdbe.org/7poy PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7poy RCSB], [https://www.ebi.ac.uk/pdbsum/7poy PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7poy ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[https://www.uniprot.org/uniprot/IPNA_EMENI IPNA_EMENI]] Isopenicillin N synthase; part of the gene cluster that mediates the biosynthesis of penicillin, the world's most important antibiotic (PubMed:3319778, PubMed:11755401). IpnA catalyzes the cyclization of the tripeptide N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine (LLD-ACV or ACV) to form isopenicillin N (IPN) that contains the beta-lactam nucleus (PubMed:3319778, PubMed:11755401, PubMed:28703303). The penicillin biosynthesis occurs via 3 enzymatic steps, the first corresponding to the production of the tripeptide N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine (LLD-ACV or ACV) by the NRPS acvA. The tripeptide ACV is then cyclized to isopenicillin N (IPN) by the isopenicillin N synthase ipnA that forms the beta-lactam nucleus. Finally, the alpha-aminoadipyl side chain is exchanged for phenylacetic acid by the isopenicillin N acyltransferase penDE to yield penicillin in the peroxisomal matrix (By similarity).[UniProtKB:P08703]<ref>PMID:11755401</ref> <ref>PMID:28703303</ref> <ref>PMID:3319778</ref>
| + | [https://www.uniprot.org/uniprot/IPNA_EMENI IPNA_EMENI] Isopenicillin N synthase; part of the gene cluster that mediates the biosynthesis of penicillin, the world's most important antibiotic (PubMed:11755401, PubMed:3319778). IpnA catalyzes the cyclization of the tripeptide N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine (LLD-ACV or ACV) to form isopenicillin N (IPN) that contains the beta-lactam nucleus (PubMed:11755401, PubMed:28703303, PubMed:3319778). The penicillin biosynthesis occurs via 3 enzymatic steps, the first corresponding to the production of the tripeptide N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine (LLD-ACV or ACV) by the NRPS acvA. The tripeptide ACV is then cyclized to isopenicillin N (IPN) by the isopenicillin N synthase ipnA that forms the beta-lactam nucleus. Finally, the alpha-aminoadipyl side chain is exchanged for phenylacetic acid by the isopenicillin N acyltransferase penDE to yield penicillin in the peroxisomal matrix (By similarity).[UniProtKB:P08703]<ref>PMID:11755401</ref> <ref>PMID:28703303</ref> <ref>PMID:3319778</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
| - | Isopenicillin N synthase (IPNS) catalyses formation of the beta-lactam and thiazolidine rings of isopenicillin N (IPN) from its linear tripeptide L-delta-(alpha-aminoadipoyl)-L-cysteinyl-D-valine (ACV) substrate in an iron and dioxygen (O2) dependent four electron oxidation without precedent in current synthetic chemistry. Recent X-ray free electron laser (XFEL) studies including time-resolved serial femtosecond crystallography show binding of O2 to the IPNS:Fe(II):ACV complex induces unexpected conformational changes in alpha-helices on the surface of IPNS, in particular in alpha3 and alpha10. However, how substrate binding leads to conformational changes away from the active site is unknown. Here, using detailed (19)F NMR and EPR experiments with labelled IPNS variants, we investigated motions in alpha3 and alpha10 induced by binding of ferrous iron, ACV and the O2 analogue NO, using the less mobile alpha6 for comparison. (19)F NMR studies were carried out on singly and doubly labelled alpha3, alpha6 and alpha10 variants at different temperatures. In addition, double electron-electron resonance (DEER) EPR analysis was carried out on doubly spin labelled variants. The combined spectroscopic and crystallographic results reveal that substantial conformational changes in regions of IPNS including alpha3 and alpha10 are induced by binding of ACV and NO. Since IPNS is a member of the structural superfamily of 2-oxoglutarate dependent oxygenases and related enzymes, related conformational changes may be of general importance in non-heme oxygenase catalysis. | + | Isopenicillin N synthase (IPNS) catalyzes formation of the beta-lactam and thiazolidine rings of isopenicillin N from its linear tripeptide l-delta-(alpha-aminoadipoyl)-l-cysteinyl-d-valine (ACV) substrate in an iron- and dioxygen (O(2))-dependent four-electron oxidation without precedent in current synthetic chemistry. Recent X-ray free-electron laser studies including time-resolved serial femtosecond crystallography show that binding of O(2) to the IPNS-Fe(II)-ACV complex induces unexpected conformational changes in alpha-helices on the surface of IPNS, in particular in alpha3 and alpha10. However, how substrate binding leads to conformational changes away from the active site is unknown. Here, using detailed (19)F NMR and electron paramagnetic resonance experiments with labeled IPNS variants, we investigated motions in alpha3 and alpha10 induced by binding of ferrous iron, ACV, and the O(2) analog nitric oxide, using the less mobile alpha6 for comparison. (19)F NMR studies were carried out on singly and doubly labeled alpha3, alpha6, and alpha10 variants at different temperatures. In addition, double electron-electron resonance electron paramagnetic resonance analysis was carried out on doubly spin-labeled variants. The combined spectroscopic and crystallographic results reveal that substantial conformational changes in regions of IPNS including alpha3 and alpha10 are induced by binding of ACV and nitric oxide. Since IPNS is a member of the structural superfamily of 2-oxoglutarate-dependent oxygenases and related enzymes, related conformational changes may be of general importance in nonheme oxygenase catalysis. |
| | | | |
| - | Spectroscopic studies reveal details of substrate-induced conformational changes distant from the active site in isopenicillin N synthase.,Rabe P, Walla CC, Goodyear NK, Welsh J, Southwart R, Clifton I, Linyard JDS, Tumber A, Claridge TDW, Myers WK, Schofield CJ J Biol Chem. 2022 Jul 11:102249. doi: 10.1016/j.jbc.2022.102249. PMID:35835215<ref>PMID:35835215</ref> | + | Spectroscopic studies reveal details of substrate-induced conformational changes distant from the active site in isopenicillin N synthase.,Rabe P, Walla CC, Goodyear NK, Welsh J, Southwart R, Clifton I, Linyard JDS, Tumber A, Claridge TDW, Myers WK, Schofield CJ J Biol Chem. 2022 Sep;298(9):102249. doi: 10.1016/j.jbc.2022.102249. Epub 2022 , Jul 11. PMID:35835215<ref>PMID:35835215</ref> |
| | | | |
| | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> |
| | </div> | | </div> |
| | <div class="pdbe-citations 7poy" style="background-color:#fffaf0;"></div> | | <div class="pdbe-citations 7poy" style="background-color:#fffaf0;"></div> |
| | + | |
| | + | ==See Also== |
| | + | *[[Isopenicillin N synthase|Isopenicillin N synthase]] |
| | == References == | | == References == |
| | <references/> | | <references/> |
| Structural highlights
7poy is a 1 chain structure with sequence from Aspergillus nidulans FGSC A4. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
| | Method: | X-ray diffraction, Resolution 1.75Å |
| Ligands: | , , , , |
| Resources: | FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT |
Function
IPNA_EMENI Isopenicillin N synthase; part of the gene cluster that mediates the biosynthesis of penicillin, the world's most important antibiotic (PubMed:11755401, PubMed:3319778). IpnA catalyzes the cyclization of the tripeptide N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine (LLD-ACV or ACV) to form isopenicillin N (IPN) that contains the beta-lactam nucleus (PubMed:11755401, PubMed:28703303, PubMed:3319778). The penicillin biosynthesis occurs via 3 enzymatic steps, the first corresponding to the production of the tripeptide N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine (LLD-ACV or ACV) by the NRPS acvA. The tripeptide ACV is then cyclized to isopenicillin N (IPN) by the isopenicillin N synthase ipnA that forms the beta-lactam nucleus. Finally, the alpha-aminoadipyl side chain is exchanged for phenylacetic acid by the isopenicillin N acyltransferase penDE to yield penicillin in the peroxisomal matrix (By similarity).[UniProtKB:P08703][1] [2] [3]
Publication Abstract from PubMed
Isopenicillin N synthase (IPNS) catalyzes formation of the beta-lactam and thiazolidine rings of isopenicillin N from its linear tripeptide l-delta-(alpha-aminoadipoyl)-l-cysteinyl-d-valine (ACV) substrate in an iron- and dioxygen (O(2))-dependent four-electron oxidation without precedent in current synthetic chemistry. Recent X-ray free-electron laser studies including time-resolved serial femtosecond crystallography show that binding of O(2) to the IPNS-Fe(II)-ACV complex induces unexpected conformational changes in alpha-helices on the surface of IPNS, in particular in alpha3 and alpha10. However, how substrate binding leads to conformational changes away from the active site is unknown. Here, using detailed (19)F NMR and electron paramagnetic resonance experiments with labeled IPNS variants, we investigated motions in alpha3 and alpha10 induced by binding of ferrous iron, ACV, and the O(2) analog nitric oxide, using the less mobile alpha6 for comparison. (19)F NMR studies were carried out on singly and doubly labeled alpha3, alpha6, and alpha10 variants at different temperatures. In addition, double electron-electron resonance electron paramagnetic resonance analysis was carried out on doubly spin-labeled variants. The combined spectroscopic and crystallographic results reveal that substantial conformational changes in regions of IPNS including alpha3 and alpha10 are induced by binding of ACV and nitric oxide. Since IPNS is a member of the structural superfamily of 2-oxoglutarate-dependent oxygenases and related enzymes, related conformational changes may be of general importance in nonheme oxygenase catalysis.
Spectroscopic studies reveal details of substrate-induced conformational changes distant from the active site in isopenicillin N synthase.,Rabe P, Walla CC, Goodyear NK, Welsh J, Southwart R, Clifton I, Linyard JDS, Tumber A, Claridge TDW, Myers WK, Schofield CJ J Biol Chem. 2022 Sep;298(9):102249. doi: 10.1016/j.jbc.2022.102249. Epub 2022 , Jul 11. PMID:35835215[4]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Ogle JM, Clifton IJ, Rutledge PJ, Elkins JM, Burzlaff NI, Adlington RM, Roach PL, Baldwin JE. Alternative oxidation by isopenicillin N synthase observed by X-ray diffraction. Chem Biol. 2001 Dec;8(12):1231-7. PMID:11755401
- ↑ McNeill LA, Brown TJN, Sami M, Clifton IJ, Burzlaff NI, Claridge TDW, Adlington RM, Baldwin JE, Rutledge PJ, Schofield CJ. Terminally Truncated Isopenicillin N Synthase Generates a Dithioester Product: Evidence for a Thioaldehyde Intermediate during Catalysis and a New Mode of Reaction for Non-Heme Iron Oxidases. Chemistry. 2017 Sep 18;23(52):12815-12824. doi: 10.1002/chem.201701592. Epub 2017, Aug 21. PMID:28703303 doi:http://dx.doi.org/10.1002/chem.201701592
- ↑ Ramon D, Carramolino L, Patino C, Sanchez F, Penalva MA. Cloning and characterization of the isopenicillin N synthetase gene mediating the formation of the beta-lactam ring in Aspergillus nidulans. Gene. 1987;57(2-3):171-81. doi: 10.1016/0378-1119(87)90120-x. PMID:3319778 doi:http://dx.doi.org/10.1016/0378-1119(87)90120-x
- ↑ Rabe P, Walla CC, Goodyear NK, Welsh J, Southwart R, Clifton I, Linyard JDS, Tumber A, Claridge TDW, Myers WK, Schofield CJ. Spectroscopic studies reveal details of substrate-induced conformational changes distant from the active site in isopenicillin N synthase. J Biol Chem. 2022 Jul 11:102249. doi: 10.1016/j.jbc.2022.102249. PMID:35835215 doi:http://dx.doi.org/10.1016/j.jbc.2022.102249
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