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| | ==Solution structure of the apo doublet acyl carrier protein from prodigiosin biosynthesis== | | ==Solution structure of the apo doublet acyl carrier protein from prodigiosin biosynthesis== |
| - | <StructureSection load='5y08' size='340' side='right'caption='[[5y08]], [[NMR_Ensembles_of_Models | 20 NMR models]]' scene=''> | + | <StructureSection load='5y08' size='340' side='right'caption='[[5y08]]' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[5y08]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/Sers3 Sers3]. Full experimental information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5Y08 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5Y08 FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[5y08]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Serratia_sp._ATCC_39006 Serratia sp. ATCC 39006]. Full experimental information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5Y08 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5Y08 FirstGlance]. <br> |
| - | </td></tr><tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">pigH ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=104623 SERS3])</td></tr> | + | </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=5y08 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5y08 OCA], [https://pdbe.org/5y08 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5y08 RCSB], [https://www.ebi.ac.uk/pdbsum/5y08 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5y08 ProSAT]</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=5y08 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5y08 OCA], [http://pdbe.org/5y08 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5y08 RCSB], [http://www.ebi.ac.uk/pdbsum/5y08 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5y08 ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/PIGH_SERS3 PIGH_SERS3]] Involved in the biosynthesis of 4-methoxy-2,2'-bipyrrole-5-carbaldehyde (MBC), one of the terminal products involved in the biosynthesis of the red antibiotic prodigiosin (Pig). Carrier of the L-malonyl group (malonyl-S-PigH), which is decarboxylated by PigJ to yield a C2 carbanion acetyl-S-PigH. Then the pyrrolyl group of pyrrolyl-S-cysteinyl PigJ intermediate is captured by the C2 carbanion acetyl-S-PigH to yield the pyrrolyl-beta-ketoacyl-S-PigH. In the last step, PigH catalyzes the decarboxylative condensation between the pyrrolyl-beta-ketoacyl (pyrrolyl-beta-ketoacyl-S-PigH) and L-serine to yield 4-hydroxy-2,2'-bipyrrole-5-methanol (HBM).<ref>PMID:15853884</ref> <ref>PMID:17002325</ref> | + | [https://www.uniprot.org/uniprot/PIGH_SERS3 PIGH_SERS3] Involved in the biosynthesis of 4-methoxy-2,2'-bipyrrole-5-carbaldehyde (MBC), one of the terminal products involved in the biosynthesis of the red antibiotic prodigiosin (Pig). Carrier of the L-malonyl group (malonyl-S-PigH), which is decarboxylated by PigJ to yield a C2 carbanion acetyl-S-PigH. Then the pyrrolyl group of pyrrolyl-S-cysteinyl PigJ intermediate is captured by the C2 carbanion acetyl-S-PigH to yield the pyrrolyl-beta-ketoacyl-S-PigH. In the last step, PigH catalyzes the decarboxylative condensation between the pyrrolyl-beta-ketoacyl (pyrrolyl-beta-ketoacyl-S-PigH) and L-serine to yield 4-hydroxy-2,2'-bipyrrole-5-methanol (HBM).<ref>PMID:15853884</ref> <ref>PMID:17002325</ref> |
| | + | <div style="background-color:#fffaf0;"> |
| | + | == Publication Abstract from PubMed == |
| | + | The acyl carrier protein (ACP) is an indispensable component of both fatty acid and polyketide synthases and is primarily responsible for delivering acyl intermediates to enzymatic partners. At present, increasing numbers of multidomain ACPs have been discovered with roles in molecular recognition of trans-acting enzymatic partners as well as increasing metabolic flux. Further structural information is required to provide insight into their function, yet to date, the only high-resolution structure of this class to be determined is that of the doublet ACP (two continuous ACP domains) from mupirocin synthase. Here we report the solution nuclear magnetic resonance (NMR) structure of the doublet ACP domains from PigH (PigH ACP(1)-ACP(2)), which is an enzyme that catalyzes the formation of the bipyrrolic intermediate of prodigiosin, a potent anticancer compound with a variety of biological activities. The PigH ACP(1)-ACP(2) structure shows each ACP domain consists of three conserved helices connected by a linker that is partially restricted by interactions with the ACP(1) domain. Analysis of the holo (4'-phosphopantetheine, 4'-PP) form of PigH ACP(1)-ACP(2) by NMR revealed conformational exchange found predominantly in the ACP(2) domain reflecting the inherent plasticity of this ACP. Furthermore, ensemble models obtained from SAXS data reveal two distinct conformers, bent and extended, of both apo (unmodified) and holo PigH ACP(1)-ACP(2) mediated by the central linker. The bent conformer appears to be a result of linker-ACP(1) interactions detected by NMR and might be important for intradomain communication during the biosynthesis. These results provide new insights into the behavior of the interdomain linker of multiple ACP domains that may modulate protein-protein interactions. This is likely to become an increasingly important consideration for metabolic engineering in prodigiosin and other related biosynthetic pathways. |
| | + | |
| | + | Solution Structure and Conformational Dynamics of a Doublet Acyl Carrier Protein from Prodigiosin Biosynthesis.,Thongkawphueak T, Winter AJ, Williams C, Maple HJ, Soontaranon S, Kaewhan C, Campopiano DJ, Crump MP, Wattana-Amorn P Biochemistry. 2021 Jan 26;60(3):219-230. doi: 10.1021/acs.biochem.0c00830. Epub , 2021 Jan 8. PMID:33416314<ref>PMID:33416314</ref> |
| | + | |
| | + | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> |
| | + | </div> |
| | + | <div class="pdbe-citations 5y08" style="background-color:#fffaf0;"></div> |
| | == References == | | == References == |
| | <references/> | | <references/> |
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| | </StructureSection> | | </StructureSection> |
| | [[Category: Large Structures]] | | [[Category: Large Structures]] |
| - | [[Category: Sers3]] | + | [[Category: Serratia sp. ATCC 39006]] |
| - | [[Category: Wattana-amorn, P]] | + | [[Category: Wattana-amorn P]] |
| - | [[Category: Acyl carrier protein]]
| + | |
| - | [[Category: Biosynthetic protein]]
| + | |
| Structural highlights
Function
PIGH_SERS3 Involved in the biosynthesis of 4-methoxy-2,2'-bipyrrole-5-carbaldehyde (MBC), one of the terminal products involved in the biosynthesis of the red antibiotic prodigiosin (Pig). Carrier of the L-malonyl group (malonyl-S-PigH), which is decarboxylated by PigJ to yield a C2 carbanion acetyl-S-PigH. Then the pyrrolyl group of pyrrolyl-S-cysteinyl PigJ intermediate is captured by the C2 carbanion acetyl-S-PigH to yield the pyrrolyl-beta-ketoacyl-S-PigH. In the last step, PigH catalyzes the decarboxylative condensation between the pyrrolyl-beta-ketoacyl (pyrrolyl-beta-ketoacyl-S-PigH) and L-serine to yield 4-hydroxy-2,2'-bipyrrole-5-methanol (HBM).[1] [2]
Publication Abstract from PubMed
The acyl carrier protein (ACP) is an indispensable component of both fatty acid and polyketide synthases and is primarily responsible for delivering acyl intermediates to enzymatic partners. At present, increasing numbers of multidomain ACPs have been discovered with roles in molecular recognition of trans-acting enzymatic partners as well as increasing metabolic flux. Further structural information is required to provide insight into their function, yet to date, the only high-resolution structure of this class to be determined is that of the doublet ACP (two continuous ACP domains) from mupirocin synthase. Here we report the solution nuclear magnetic resonance (NMR) structure of the doublet ACP domains from PigH (PigH ACP(1)-ACP(2)), which is an enzyme that catalyzes the formation of the bipyrrolic intermediate of prodigiosin, a potent anticancer compound with a variety of biological activities. The PigH ACP(1)-ACP(2) structure shows each ACP domain consists of three conserved helices connected by a linker that is partially restricted by interactions with the ACP(1) domain. Analysis of the holo (4'-phosphopantetheine, 4'-PP) form of PigH ACP(1)-ACP(2) by NMR revealed conformational exchange found predominantly in the ACP(2) domain reflecting the inherent plasticity of this ACP. Furthermore, ensemble models obtained from SAXS data reveal two distinct conformers, bent and extended, of both apo (unmodified) and holo PigH ACP(1)-ACP(2) mediated by the central linker. The bent conformer appears to be a result of linker-ACP(1) interactions detected by NMR and might be important for intradomain communication during the biosynthesis. These results provide new insights into the behavior of the interdomain linker of multiple ACP domains that may modulate protein-protein interactions. This is likely to become an increasingly important consideration for metabolic engineering in prodigiosin and other related biosynthetic pathways.
Solution Structure and Conformational Dynamics of a Doublet Acyl Carrier Protein from Prodigiosin Biosynthesis.,Thongkawphueak T, Winter AJ, Williams C, Maple HJ, Soontaranon S, Kaewhan C, Campopiano DJ, Crump MP, Wattana-Amorn P Biochemistry. 2021 Jan 26;60(3):219-230. doi: 10.1021/acs.biochem.0c00830. Epub , 2021 Jan 8. PMID:33416314[3]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Williamson NR, Simonsen HT, Ahmed RA, Goldet G, Slater H, Woodley L, Leeper FJ, Salmond GP. Biosynthesis of the red antibiotic, prodigiosin, in Serratia: identification of a novel 2-methyl-3-n-amyl-pyrrole (MAP) assembly pathway, definition of the terminal condensing enzyme, and implications for undecylprodigiosin biosynthesis in Streptomyces. Mol Microbiol. 2005 May;56(4):971-89. PMID:15853884 doi:http://dx.doi.org/10.1111/j.1365-2958.2005.04602.x
- ↑ Garneau-Tsodikova S, Dorrestein PC, Kelleher NL, Walsh CT. Protein assembly line components in prodigiosin biosynthesis: characterization of PigA,G,H,I,J. J Am Chem Soc. 2006 Oct 4;128(39):12600-1. PMID:17002325 doi:http://dx.doi.org/10.1021/ja063611l
- ↑ Thongkawphueak T, Winter AJ, Williams C, Maple HJ, Soontaranon S, Kaewhan C, Campopiano DJ, Crump MP, Wattana-Amorn P. Solution Structure and Conformational Dynamics of a Doublet Acyl Carrier Protein from Prodigiosin Biosynthesis. Biochemistry. 2021 Jan 26;60(3):219-230. PMID:33416314 doi:10.1021/acs.biochem.0c00830
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