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| Line 3: |
Line 3: |
| | <StructureSection load='7s0l' size='340' side='right'caption='[[7s0l]], [[Resolution|resolution]] 2.65Å' scene=''> | | <StructureSection load='7s0l' size='340' side='right'caption='[[7s0l]], [[Resolution|resolution]] 2.65Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[7s0l]] is a 2 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7S0L OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7S0L FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[7s0l]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Talaromyces_verruculosus Talaromyces verruculosus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7S0L OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7S0L FirstGlance]. <br> |
| - | </td></tr><tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat"><div style='overflow: auto; max-height: 3em;'>[[6v0k|6v0k]]</div></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]] 2.65Å</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=7s0l FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7s0l OCA], [https://pdbe.org/7s0l PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7s0l RCSB], [https://www.ebi.ac.uk/pdbsum/7s0l PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7s0l 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=7s0l FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7s0l OCA], [https://pdbe.org/7s0l PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7s0l RCSB], [https://www.ebi.ac.uk/pdbsum/7s0l PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7s0l ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/PVCPS_TALVE PVCPS_TALVE] Bifunctional terpene synthase that possesses both prenyltransferase and type II terpene cyclase activity, converting isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) into geranylgeranyl diphosphate (GGPP) and further converting GGPP into copalyl diphosphate, respectively.<ref>PMID:28869716</ref> <ref>PMID:31978464</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
| Line 21: |
Line 23: |
| | </StructureSection> | | </StructureSection> |
| | [[Category: Large Structures]] | | [[Category: Large Structures]] |
| - | [[Category: Christianson, D W]] | + | [[Category: Talaromyces verruculosus]] |
| - | [[Category: Ronnebaum, T A]] | + | [[Category: Christianson DW]] |
| - | [[Category: Assembly-line synthase]] | + | [[Category: Ronnebaum TA]] |
| - | [[Category: Bifunctional terpene synthase]]
| + | |
| - | [[Category: Ggpp synthase]]
| + | |
| - | [[Category: Isoprenoid synthase]]
| + | |
| - | [[Category: Prenyltransferase]]
| + | |
| - | [[Category: Transferase]]
| + | |
| Structural highlights
Function
PVCPS_TALVE Bifunctional terpene synthase that possesses both prenyltransferase and type II terpene cyclase activity, converting isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) into geranylgeranyl diphosphate (GGPP) and further converting GGPP into copalyl diphosphate, respectively.[1] [2]
Publication Abstract from PubMed
Copalyl diphosphate (CPP) synthase from Penicillium verruculosum (PvCPS) is a bifunctional diterpene synthase with both prenyltransferase and class II cyclase activities. The prenyltransferase alpha domain catalyzes the condensation of C5 dimethylallyl diphosphate with three successively added C5 isopentenyl diphosphates (IPPs) to form C20 geranylgeranyl diphosphate (GGPP), which then undergoes a class II cyclization reaction at the betagamma domain interface to generate CPP. The prenyltransferase alpha domain mediates oligomerization to form a 648-kD (alphabetagamma)6 hexamer. In the current study, we explore prenyltransferase structure-function relationships in this oligomeric assembly-line platform with the goal of generating alternative linear isoprenoid products. Specifically, we report steady-state enzyme kinetics, product analysis, and crystal structures of various site-specific variants of the prenyltransferase alpha domain. Crystal structures of the H786A, F760A, S723Y, S723F, and S723T variants have been determined at resolutions of 2.80, 3.10, 3.15, 2.65, and 2.00 A, respectively. The substitution of S723 with bulky aromatic amino acids in the S723Y and S723F variants constricts the active site, thereby directing the formation of the shorter C15 isoprenoid, farnesyl diphosphate. While the S723T substitution only subtly alters enzyme kinetics and does not compromise GGPP biosynthesis, the crystal structure of this variant reveals a nonproductive binding mode for IPP that likely accounts for substrate inhibition at high concentrations. Finally, mutagenesis of the catalytic general acid in the class II cyclase domain, D313A, significantly compromises prenyltransferase activity. This result suggests molecular communication between the prenyltransferase and cyclase domains despite their distant connection by a flexible polypeptide linker.
Engineering the Prenyltransferase Domain of a Bifunctional Assembly-Line Terpene Synthase.,Ronnebaum TA, Eaton SA, Brackhahn EAE, Christianson DW Biochemistry. 2021 Oct 5. doi: 10.1021/acs.biochem.1c00600. PMID:34609847[3]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Mitsuhashi T, Okada M, Abe I. Identification of Chimeric αβγ Diterpene Synthases Possessing both Type II Terpene Cyclase and Prenyltransferase Activities. Chembiochem. 2017 Nov 2;18(21):2104-2109. PMID:28869716 doi:10.1002/cbic.201700445
- ↑ Ronnebaum TA, Gupta K, Christianson DW. Higher-Order Oligomerization of a Chimeric alphabetagamma Bifunctional Diterpene Synthase with Prenyltransferase and Class II Cyclase Activities is Concentration-Dependent. J Struct Biol. 2020 Jan 21:107463. doi: 10.1016/j.jsb.2020.107463. PMID:31978464 doi:http://dx.doi.org/10.1016/j.jsb.2020.107463
- ↑ Ronnebaum TA, Eaton SA, Brackhahn EAE, Christianson DW. Engineering the Prenyltransferase Domain of a Bifunctional Assembly-Line Terpene Synthase. Biochemistry. 2021 Oct 5. doi: 10.1021/acs.biochem.1c00600. PMID:34609847 doi:http://dx.doi.org/10.1021/acs.biochem.1c00600
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