Combinatorial Assembly of Simple and Complex D-Lysergic Acid Alkaloid Peptide Classes in the Ergot Fungus Claviceps purpurea
Combinatorial Assembly of Simple and Complex D-Lysergic Acid Alkaloid Peptide Classes in the Ergot Fungus Claviceps purpurea
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DOI：
10.1074/jbc.m807168200
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发表时间：
2009-03-13
影响因子：
4.8
通讯作者：
Keller, Ullrich
中科院分区：
文献类型：
--
作者：
Ortel, Ingo;Keller, Ullrich
麦角菌麦角菌产生麦角肽和简单的D-麦角酸烷基酰胺。在麦角肽（例如麦角胺）中，D-麦角酸以类似酰胺的方式与双环三肽连接，而在D-麦角基烷醇酰胺中，它与源自丙氨酸的氨基醇连接。在这里，我们表明，这些化合物类合成的一组三个非核糖体麦角酰肽合成酶（LPS），相互作用的组合方式合成的相关产品。三模块LPS 1与LPS 2（D-麦角酸募集模块）组装，以从D-麦角酸和肽链的三个氨基酸合成麦角肽的D-麦角酰三肽前体。或者，LPS 2可以与独特的单模非核糖体肽合成酶（NRPS）亚基（麦角新碱合成酶）组装，以从D-麦角酸和丙氨酸合成D-麦角酸烷醇酰胺麦角新碱。合成通过共价结合的D-麦角酰丙氨酸和二肽作为醇释放并消耗NADPH来进行。酶和免疫化学分析表明麦角新碱合成酶可能是LPS 3，其基因已被确定为麦角生物碱合成基因簇的一部分。紫花。所有LPS序列的检查表明，没有可识别的肽接头的蛋白质-蛋白质相互作用的NRPS亚基的细菌。相反，它们都携带保守的N-末端结构域（C 0-结构域），与NRPS缩合结构域的C-末端半部分相似，指向真菌NRPS系统中亚基-亚基相互作用的替代机制。LPS模块和C 0-结构域的系统发育分析表明，这些酶系统最有可能从一个双模祖先的模块复制和重排演变。
The ergot fungus Claviceps purpurea produces both ergopeptines and simple D-lysergic acid alkylamides. In the ergopeptines, such as ergotamine, D-lysergic acid is linked to a bicyclic tripeptide in amide-like fashion, whereas in the D-lysergylalkanolamides it is linked to an amino alcohol derived from alanine. We show here that these compound classes are synthesized by a set of three non-ribosomal lysergyl peptide synthetases (LPSs), which interact in a combinatorial fashion for synthesis of the relevant product. The trimodular LPS1 assembles with LPS2, the D-lysergic acid recruiting module, to synthesize the D-lysergyltripeptide precursors of ergopeptines from D-lysergic acid and the three amino acids of the peptide chain. Alternatively, LPS2 can assemble with a distinct monomodular non-ribosomal peptide synthetase (NRPS) subunit (ergometrine synthetase) to synthesize the D-lysergic acid alkanolamide ergometrine from D-lysergic acid and alanine. The synthesis proceeds via covalently bound D-lysergyl alanine and release of dipeptide as alcohol with consumption of NADPH. Enzymatic and immunochemical analyses showed that ergometrine synthetase is most probably the enzyme LPS3 whose gene had been identified previously as part of the ergot alkaloid biosynthesis gene cluster in C. purpurea. Inspections of all LPS sequences showed no recognizable peptide linkers for their protein-protein interactions as in NRPS subunits of bacteria. Instead, they all carry conserved N-terminal domains (C0-domains) with similarity to the C-terminal halves of NRPS condensation domains pointing to an alternative mechanism of subunit-subunit interactions in fungal NRPS systems. Phylogenetic analysis of LPS modules and the C0-domains suggests that these enzyme systems most probably evolved by module duplications and rearrangements from a bimodular ancestor.