4nao

Structural highlights

Function

EASH_CLAP2 Dioxygenase; part of the gene cluster that mediates the biosynthesis of fungal ergot alkaloid (PubMed:14732265, PubMed:14700635, PubMed:15904941, PubMed:17308187, PubMed:17720822). DmaW catalyzes the first step of ergot alkaloid biosynthesis by condensing dimethylallyl diphosphate (DMAP) and tryptophan to form 4-dimethylallyl-L-tryptophan (PubMed:14732265). The second step is catalyzed by the methyltransferase easF that methylates 4-dimethylallyl-L-tryptophan in the presence of S-adenosyl-L-methionine, resulting in the formation of 4-dimethylallyl-L-abrine (By similarity). The catalase easC and the FAD-dependent oxidoreductase easE then transform 4-dimethylallyl-L-abrine to chanoclavine-I which is further oxidized by easD in the presence of NAD(+), resulting in the formation of chanoclavine-I aldehyde (PubMed:20118373, PubMed:21409592). Agroclavine dehydrogenase easG then mediates the conversion of chanoclavine-I aldehyde to agroclavine via a non-enzymatic adduct reaction: the substrate is an iminium intermediate that is formed spontaneously from chanoclavine-I aldehyde in the presence of glutathione (PubMed:20735127, PubMed:21494745). The presence of easA is not required to complete this reaction (PubMed:21494745). Further conversion of agroclavine to paspalic acid is a two-step process involving oxidation of agroclavine to elymoclavine and of elymoclavine to paspalic acid, the second step being performed by the elymoclavine oxidase cloA (PubMed:16538694, PubMed:17720822). Paspalic acid is then further converted to D-lysergic acid (PubMed:15904941). Ergopeptines are assembled from D-lysergic acid and three different amino acids by the D-lysergyl-peptide-synthetases composed each of a monomudular and a trimodular nonribosomal peptide synthetase subunit (PubMed:14700635, PubMed:15904941). LpsB and lpsC encode the monomodular subunits responsible for D-lysergic acid activation and incorporation into the ergopeptine backbone (PubMed:14700635). LpsA1 and A2 subunits encode the trimodular nonribosomal peptide synthetase assembling the tripeptide portion of ergopeptines (PubMed:14700635). LpsA1 is responsible for formation of the major ergopeptine, ergotamine, and lpsA2 for alpha-ergocryptine, the minor ergopeptine of the total alkaloid mixture elaborated by C.purpurea (PubMed:17560817, PubMed:19139103). D-lysergyl-tripeptides are assembled by the nonribosomal peptide synthetases and released as N-(D-lysergyl-aminoacyl)-lactams (PubMed:24361048). Cyclolization of the D-lysergyl-tripeptides is performed by the Fe(2+)/2-ketoglutarate-dependent dioxygenase easH which introduces a hydroxyl group into N-(D-lysergyl-aminoacyl)-lactam at alpha-C of the aminoacyl residue followed by spontaneous condensation with the terminal lactam carbonyl group (PubMed:24361048).[UniProtKB:Q50EL0]^{[1] [2] [3] [4] [5] [6] [7] [8] [9] [10] [11] [12] [13]}

References

1. ↑ Correia T, Grammel N, Ortel I, Keller U, Tudzynski P. Molecular cloning and analysis of the ergopeptine assembly system in the ergot fungus Claviceps purpurea. Chem Biol. 2003 Dec;10(12):1281-92. doi: 10.1016/j.chembiol.2003.11.013. PMID:14700635 doi:http://dx.doi.org/10.1016/j.chembiol.2003.11.013
2. ↑ Wang J, Machado C, Panaccione DG, Tsai HF, Schardl CL. The determinant step in ergot alkaloid biosynthesis by an endophyte of perennial ryegrass. Fungal Genet Biol. 2004 Feb;41(2):189-98. PMID:14732265
3. ↑ Haarmann T, Machado C, Lubbe Y, Correia T, Schardl CL, Panaccione DG, Tudzynski P. The ergot alkaloid gene cluster in Claviceps purpurea: extension of the cluster sequence and intra species evolution. Phytochemistry. 2005 Jun;66(11):1312-20. doi: 10.1016/j.phytochem.2005.04.011. PMID:15904941 doi:http://dx.doi.org/10.1016/j.phytochem.2005.04.011
4. ↑ Haarmann T, Ortel I, Tudzynski P, Keller U. Identification of the cytochrome P450 monooxygenase that bridges the clavine and ergoline alkaloid pathways. Chembiochem. 2006 Apr;7(4):645-52. doi: 10.1002/cbic.200500487. PMID:16538694 doi:http://dx.doi.org/10.1002/cbic.200500487
5. ↑ Haarmann T, Lorenz N, Tudzynski P. Use of a nonhomologous end joining deficient strain (Deltaku70) of the ergot fungus Claviceps purpurea for identification of a nonribosomal peptide synthetase gene involved in ergotamine biosynthesis. Fungal Genet Biol. 2008 Jan;45(1):35-44. doi: 10.1016/j.fgb.2007.04.008. Epub , 2007 May 10. PMID:17560817 doi:http://dx.doi.org/10.1016/j.fgb.2007.04.008
6. ↑ Ortel I, Keller U. Combinatorial assembly of simple and complex D-lysergic acid alkaloid peptide classes in the ergot fungus Claviceps purpurea. J Biol Chem. 2009 Mar 13;284(11):6650-60. doi: 10.1074/jbc.M807168200. Epub 2009 , Jan 12. PMID:19139103 doi:http://dx.doi.org/10.1074/jbc.M807168200
7. ↑ Lorenz N, Olsovska J, Sulc M, Tudzynski P. Alkaloid cluster gene ccsA of the ergot fungus Claviceps purpurea encodes chanoclavine I synthase, a flavin adenine dinucleotide-containing oxidoreductase mediating the transformation of N-methyl-dimethylallyltryptophan to chanoclavine I. Appl Environ Microbiol. 2010 Mar;76(6):1822-30. doi: 10.1128/AEM.00737-09. Epub , 2010 Jan 29. PMID:20118373 doi:http://dx.doi.org/10.1128/AEM.00737-09
8. ↑ Cheng JZ, Coyle CM, Panaccione DG, O'Connor SE. Controlling a structural branch point in ergot alkaloid biosynthesis. J Am Chem Soc. 2010 Sep 22;132(37):12835-7. doi: 10.1021/ja105785p. PMID:20735127 doi:http://dx.doi.org/10.1021/ja105785p
9. ↑ Goetz KE, Coyle CM, Cheng JZ, O'Connor SE, Panaccione DG. Ergot cluster-encoded catalase is required for synthesis of chanoclavine-I in Aspergillus fumigatus. Curr Genet. 2011 Jun;57(3):201-11. doi: 10.1007/s00294-011-0336-4. Epub 2011 Mar , 17. PMID:21409592 doi:http://dx.doi.org/10.1007/s00294-011-0336-4
10. ↑ Matuschek M, Wallwey C, Xie X, Li SM. New insights into ergot alkaloid biosynthesis in Claviceps purpurea: an agroclavine synthase EasG catalyses, via a non-enzymatic adduct with reduced glutathione, the conversion of chanoclavine-I aldehyde to agroclavine. Org Biomol Chem. 2011 Jun 7;9(11):4328-35. doi: 10.1039/c0ob01215g. Epub 2011 Apr , 15. PMID:21494745 doi:http://dx.doi.org/10.1039/c0ob01215g
11. ↑ Havemann J, Vogel D, Loll B, Keller U. Cyclolization of D-Lysergic Acid Alkaloid Peptides. Chem Biol. 2013 Dec 18. pii: S1074-5521(13)00417-1. doi:, 10.1016/j.chembiol.2013.11.008. PMID:24361048 doi:http://dx.doi.org/10.1016/j.chembiol.2013.11.008
12. ↑ Fleetwood DJ, Scott B, Lane GA, Tanaka A, Johnson RD. A complex ergovaline gene cluster in epichloe endophytes of grasses. Appl Environ Microbiol. 2007 Apr;73(8):2571-9. doi: 10.1128/AEM.00257-07. Epub , 2007 Feb 16. PMID:17308187 doi:http://dx.doi.org/10.1128/AEM.00257-07
13. ↑ Lorenz N, Wilson EV, Machado C, Schardl CL, Tudzynski P. Comparison of ergot alkaloid biosynthesis gene clusters in Claviceps species indicates loss of late pathway steps in evolution of C. fusiformis. Appl Environ Microbiol. 2007 Nov;73(22):7185-91. Epub 2007 Aug 24. PMID:17720822 doi:http://dx.doi.org/AEM.01040-07